Protein Extraction Reagent Functional Proteomics

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Protein preparation

Protein Extraction Reagent

Contains selected detergents and protease inhibitors :

• No appendix treatment required • No freezing or thawing • Short protocol : 20-30 minutes. • Get stable and non-degradated proteins • Good yields • No interference for subsequent protein absorbance measures • Ideal for protein molecular weight analysis

Name Cat# Qty

Protein Extraction Buffer (Cell/Tissue) BZ2171 100ml

UptitipTM Titanium

Phosphopeptides purification and enrichment

In the field of HPLC, it is reported that the columns packed with Titanium Dioxide (TiO2) par-ticles was applied for the analysis of phospho-compounds. UpiTip™ TiO is enable to capture selectively phosphopeptides from a comparatively large background of unmodified peptides. UptiTip™ TiO is a highly efficient and versatile tool to purify phosphorylated peptides from proteolytic digests prior mass analysis.

Feature :

• Faster sample preparation • Low sample loss • High selectivity

Product Binding Capacity

Amount of chromatographic material

Cat.# Qty/pack

UptiTip (1-10 µl) 400 micrograms 4 mg BT3530 96 uUptiTip (10-200 µl) 1000 micrograms 10 mg BU3630 96 uUptiTip (100-1000 µl) 7000 micrograms 75 mg CA5100 20 u

Proteomics - Biochemistry

Proteomics - Biochemistry of proteins

Protein Preparation

Technical tip – protein extraction procedures Procedures are proposed to 1/disrupt cells, 2/solubilize/ stabilize proteins :

• mechanical procedures : Freeze/Thawing French press, ... • detergents : choice according CMC/protein type, >see our selection guide page B2 ionic charge/downstream application • protectants : protease inhibitors, anti-oxidants, ...

Recommended volume of Protein Extraction Reagent, and typical protein yield :

Eukaryotic cells

400µl per 5.106 cells 2mg protein

Tissues 600µl per 10g 8-10mg protein


Functional Proteomics

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Proteomics - Biochemistry

IPeX Ab-free Ag Immunoprecipitation KitLeave your Ab coupled to the Immunoprecipitation support to get clean immunoblots !

Applications :

• Immunoprecipitations. • Co-immunoprecipitations. • Small-scale purifications of protein. • Small-scale purifications of recombinant protein (e.g. with His or GST tags) • Studying of protein-protein interaction. • Eliminating interference from antibody heavy and light chain bands on SDS-PAGE or

Western blots.

IPeX ImmunoPrecipitation Kit use a crosslinking method to fix covalently your primary antibody to the beads. The following immunoprecipitation yields antigens devoid of any immunoglo-bulin fragment, facilitating greatly their downstream analysis (i.e. no non-specific bands in by immunoblotting).

Kit Advantages over classical IP methods

Advantages in using an IPeX kit rather than classical IP methods :

• The affinity support can be reused up to 10 times. • Immobilizing the antibody provides faster and easier IP's. • Coupling of all primary amine-containing molecules. • Coupling of all antibody species and subclasses. (# of protein A/G methods) • Antibody is coupled directly to the beads without using a cross-linker. • Purified antigen free from antibody contamination. • Avoids on Western Blots the undesired bands due to IgGs or their heavy end Light

chains. • Short protocol

Name Cat# Qty

IPeX kit (for 10 columns preparation)Contains 10 IPeX spin columns, 10 collection tubes, beads, buffers and handbook

BI4211 Kit/10 preps

IPeX refill kit (for 100 purifications) Contains 20 ml of Binding buffer, 350 ml of Washing buffer and 20 ml Elution Buffer

BI422A Kit/100 purif.

Gentle Elution Buffer BI422h 20 ml

Also available : standard immunoprecipitation kit ImmunoCatcher, with lysis buffer :

Name Cat# Qty

IMMUNOcatcher™ (classic ImmunoPrecipitation)*Contains : 50 centrifuge filters, 100 collectors, 0.6 ml Immobilized protein A/G, 60 ml of strong lysis solution, 0.6 ml of preimmune serum, 2 ml of SDS-PAGE sample solution, Protease inhi-bitor

C02-020C02-050

20 assays50 assays*

Protein preparation

Schematic IPeX Protocol

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Proteomics - BiochemistryProtein modifications

FeBABE

Applications :

• Elucidation of 3 dimensional structures of protein complexes • Determination of protein to protein interactions • Can be also used as a chemical nuclease

This method was successfully utilized to resolve the 3D structure of many proteins, protein-protein and protein-nucleic acid complexes have been elucidated, including E.Coli cytochrome bd quinol oxidase, and a subunit of E.Coli polymerase. FeBABE allowed to identify the contact sites between many E. coli transcription factors and RNA polymerase3, to identify ribosome protein binding sites on rRNAs1, and to determine the cleavage site of the DNA or RNA.

1) G. M. Heilek, R. Marusak, C. F. Meares, and H. F. Noller. Proc. Natl. Acad. Sci. USA, 1995, 92, 1113. 2) K. R. Lieberman, and H. F. Noller, J. Mol. Biol. 1998, 284, 1367 3) S. L. Traviglia, et al,. Biochemistry, 1999, 38,15744.

Please inquire for an application technical notice.Cleavage of nucleotides and proteins by FeBABE In the presence of hydrogen peroxide and ascorbic acid, radicals are generated, which cut the DNA or pro-tein around FeBABE. By the analysis of the cleavage sites, the point-of-contact sites can be estimated.

Other special modifiers/crosslinkers for 3D structure studies

Name Cat# Qty

Iminothiolane (Traut’s reagent)Converts NH2 en SH groups

UP42425A 500mg

BNPS-Skatole Cleaves Try residues

UP20955A 1 g

MTSEA Useful in the membrane proteins mapping, as ion channels and proteins transport, as well as enzymes and receptors

UP99618 100 mg

BMOEShorter spacer (9A) amine bi-reactive

L7730A 100 mg

TCEP non-pungent and mild alternative to DTTDo not reduce buried sulfhydryl

UP242214 1 g

HPGmodify Arg residues at pK7-9

UP36862A 100mg

APGPhotoreactive and arginine selective

UP28071A 100 mg

HBVSSH reactive, but unlike maleimides, does not generate stereo isomers

UPL7733A 50 mg

Name Cat# Qty

FeBABE UP994760 1 mg

A unique tool to study the three-dimensional of proteins and interactions !

Protein modifications





ControlledAmineTM conjugation kit

The new technology to conjugate simply and efficiently your biomolecules

This technique uses hydrazone chemistry to replaces advantageously the standard methods based on NHS/Maleimide, epoxides, glutaraldehyde, ... It is more flexible and satisfying for most applications.

Advantages :

• ease of operating (modify each molecule, mix), • efficient and flexible, • activate biomolecules in advance (stable for months), • better control of the coupling ratio, • excellent yield of conjugation, • no reduction or deprotection step required, • highly selective heteroconjugation (oriented), • very stable conjugation, • keep the bioactivity of the components, • very flexible for many applications / samples.

Applications :

• label/conjugate proteins* • immobilisation on supports* • amenable to solid phase synthesis

Name Cat# Qty

Hydralink™ ControlledAmine™ Conjugation kit 1-SANHContains all reagents to conjugate 2 proteins in oriented manner

BL1501 1 kit

For detailed description of the hydrazone chemistry and its advantages, please see page B223 of the BioSciences Innovation catalogue or inquire at [email protected].

* suits any biomolecule or support containing amines or derivatized by conventional biochemistry (proteins, peptides, oligonucleo-tides, cDNAs, carbohydrates, fluorophores, beads, glass, silica ...).

Schematic IPeX Protocol

Chromalink-Biotin

A unique biotinylation agent that allows direct measure of coupled biotin

• Amine reactive at pH 7.0-9.0 • PEO absorbing at 354 nm spacer • Spectrophotometric quantitation of total biotin incorporation • Preserves biotin/avidin affinity as well as increases solubility

Single reagent Biotinylation kit

Biotin Chromalink BT3601, 5 x 0.5 mg BT3611, Kit1*BT3602, 5 x 1 mg BT3612, Kit2*

* Contains chromalink, DMF, 10X modification buffer, and 5K MWCO diafilter.

Incubation ratio HABA ratio Abs 354 ratio

5X 1.03 2.4510X 1.60 4.7115X 2.22 6.25

Bovine IgG (bIgG) reacted with different biotin ratios and subsequent determination of cou-pled ratio by Chromalink method and HABA method. The streptavidin binding efficiency of ChromaLink Biotin coupled protein was proved to be similar to Biotin-PEO4-NHS labeled protein (not shown).

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More Crosslinker and biotinylation reagents in the BioScience catalogue pages B11-B50, including > 40 PEO crosslinking agents and > 18PEO biotinylation agents

PEO/PEG Biochemistry : new reagents

PolyEthyleneGlycol (PEG) (or PolyEthylOxy : PEO) structure improves features of your conju-gates compared to conventional spacers (i.e. alkyls based) thanks :

PEG/PEO technology benefits :

• Increases water-solubility • Minimizes aggregation of conjugates or conjugates/ligands complexes • Ajustable length of spacer • No immunogenicity • Increases bio-stability • Reduces non-specific binding on surfaces

PEO features are especially taken to good account in proteomics studies, minimizing alterations of the properties of conju-gate components, and generally favoring conjugate interactions thanks superior hydrophilicity.

Spacer Functional group(s)Name Cat# Qty Lenght group 1 group 2

CrosslinkersMAL-PEO4-NHS AL6580 100mg 24.8 Ang. NHS MALMAL-PEO8-NHS BH9851 100mg 39.2 Ang. NHS MALMAL-PEO12-NHS BH9861 100mg 53.3 Ang. NHS MALMAL-PEO3-MAL(BM[PEO]3) L7735A 100mg 14.7 Ang. MAL MALMAL-PEO4-MAL(BM[PEO]4) L7736A 100mg 17.8 Ang. MAL MAL

Biotinylation agentsBiotin-PEO4-Amine 77872A 100mg 22.9 Ang. NH2 BiotinBiotin-PEO4-Hydrazide BJ008A 50mg 20.6 Ang. HYD BiotinBiotin-PEO4-Maleimide UPR2028A 25mg 38 Ang. MAL BiotinBiotin-PEO4-TFPA BT3621 10mg 33 Ang. TFPA BiotinBiotin-PEO4-NHS UPR20279

UPR2027A4x5mg50mg

NHS Biotin

Biotin-PEO12-NHS BZ0971 25mg NHS BiotinBiotin-PEO4-Maleimide UPAK789A 50mg 24.9 Ang. MAL BiotinPsoralen-PEO4-Biotin UPL77845 10mg 36.9 Ang. Psoralen BiotinBiotin-G-PEO4-COOH BJ007A 50mg 19.2Ang. COOH Biotin

MAL-PEO8-NHS

Please inquire for bulk quantities as well for manufacturing applications !

See also above our anti phosphoprotein antibodies, and below the reagents for electropho-resis / general use.

Phosphoprotein Stain KitDirect in-gel staining of proteins phosphorylated at serine and threonine residues

Phosphoproteins analysis

The Phosphoprotein Stain Kit provides direct in-gel detection of proteins phosphorylated on serine and threonine residues. The resolved phosphoproteins are visualized as green to green-blue bands. The procedure involves a hydrolysis step and a staining with Methyl Green dye. Gels can be stained with Coomassie® Blue (CooBlue, see below) after phosphoprotein staining to determine total protein composition. Each kit contains sufficient reagents to stain 5 to 10 mini-gels.

Cat # Qty

Phosphoprotein Stain Kit CD6040 1 kitIncludes : Fixing Solution 500 ml, Calcium Solution 250 ml, Hydrolysis Solution 250 ml, Amino Acid Staining Solution 250 ml, Complex Forming Solution 250 ml, Basic Dye Solution 250 ml, Destaining Solution 250 ml, Phosphoprotein Control Standards (Positive Control/Casein, 19-25 kDa and Negative Control/BSA, 66 kDa)





Here is a selection of standard fluorescent labels from our range of >200 agents (pages B51-B85 of the BioScience catalogue) + improved agents (ý), and labeling kits (©).

• Great prices • Highest quality • Large choice • Improved versions • Great Fluoprbes alternatives

Standard and improved fluorescent agents and kits

Please refer to the BioScience catalogue for a description of the labels and reactivities, or ask at [email protected].

NHS : mine reactivityMaleimide : Sulfhydryl reactivityX : extended spacer for greater bioavailabilityPEO : extended and hydrophilic spacer for higher coupling ratio and detection sensitivity

You want to get ride of limitations of standard fluorophores and expensive commercial alternatives ?

Fluorophore-linker-(Abs./em.) Qty Cat #

AMCA -ý AMCA-X-NHS (353/442nm) 10 mg FP-84695Aý FluoProbes®390A-NHS (390/479nm) 1 mg FP-BS5910

FITC (494/517nm) 100 mg FP-01739K1 g FP-01739L

ý Fluorescein-X-NHS (494/517nm) 1 mg FP-AX-1760ý FluoProbes®488-NHS (496/519nm) 1 mg FP-BA6800ý FluoProbes®488-Maleimide 1 mg FP-BA6810ý HEX-NHS (524/538nm) 10 mg FP-AM574A

TAMRA-NHS (540/565nm) 10 mg FP-52498Aý TAMRA-X-NHS (540/565nm) 10 mg FP-33406A

TRITC (543/572nm) 10 mg FP-47004Aý FluoProbes®547H-NHS (547/574nm) 1 mg FP-BX8920ý FluoProbes®547H-Maleimide 1 mg FP-CB100

SR110-SC (TR) (583/603nm) 10 mg FP-47006Aý SR110-PEO-NHS (hydrosoluble) 5 mg FP-AM409Aý FluoProbes590A (594/624nm) 5 mg FP-BA7080

Try our FluoProbes ! They are available as activated NHS, Maleimide and hydrazide deriva-tives, and in convenient labeling kits.

Fluorophore-labeling kits Cat# Qty

FluoProbes®390A Protein labeling kit 390/479 nm (compatible with standard filters for AMCA)

FP-CG5390 1 kit (5 lab.*)

FluoProbes®FITC-X Protein labeling kit 593/517 nm (Fluorescein with an extended spacer for improved fluorescent properties)

FP-AX1350 1 kit (5 lab.*)

FluoProbes®488 Protein labeling kit594/519 nm (compatible with standard filters for FITC, Cy™2)

FP-BE3750 1 kit (5 lab.*)

FluoProbes®547H Protein labeling kit 557/574 nm (compatible with standard filters for Tr, Cy™3)

FP-BZ9600 1 kit (5 lab.*)

FluoProbes®647H Protein labeling kit 652/673 nm (compatible with standard filters for Cy™5)

FP-BZ9610 1 kit (5 lab.*)

FluoProbes®682 Protein labeling kit 690/709 nm (compatible with standard filters for Cy™5.5)

FP-BE8280 1 kit (5 lab.*)

FluoProbes®782 Protein labeling kit 782/800 nm (compatible with standard filters for Cy™7)

FP-CA6070 1 kit (5 lab.*)

Kits for other FluoProbes labels are available on inquire. Labeling Kits are also available for conventional fluorophores, with convenient spin columns, NH2 and SH reactivity: see our BioScience catalogue page B61 for FITC, page B59 for PE, APC, page B89-B90 for HRP and AP enzymes.

* each labeling suit for up 1mg protein

Fluorescence emission spectra of selected fluoprobes dyes

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Numerous native proteins contain glycosidic (sugars) post-transcriptional modifications whose structures are dependent both on species and cell type. The characterization of the complex oligosaccharides obtained from these glycoproteins has proven a difficult and time consuming endeavor.

The Carbohydrate Analysis/Detection Kit allows quick estimation and/or comparison of carbohydrates samples composition. Applications include carbohydrate receptor analysis, and enzyme inhibition studies. The kit provides reagents and protocols for analyzing these carbohydrates through covalent labeling with a fluorescent reagent and analysis by thin-layer chromatography (TLC). The principle involves enzymatic removal of the oligosaccharides from a native protein (or mixture of reducing sugars), reductive amination of the reducing sugars and analysis of the resultant glycamines using silica-gel two dimensional thin layer chroma-tography (2D-TLC) or by other well established techniques (HPLC detection and purification, PAGE electrophoresis).

The advantages of the used fluorophore include its low detection limit, water solubility, pH fluorescence invariance, stability, distinctive fluorescence from protein chromophores, and ability to be detected using normal phase chromatography techniques. Additionnaly, compared with the commonly used ANTS reagent, 1/ the more nucleophilic primary amine of the dye makes it more reactive in the labeling reaction than the aromatic amine of ANTS. 2/ the dye is less polar than ANTS, having only one charged sulfate group instead of three, allowing wider potential application for a variety of carbohydrate sizes.

Glycoprotein or sugar fluorescent labeling and analysis

Product Cat# Qty

Carbohydrate Analysis/Detection Kit Contains : Fluorescent reagent, 2ml Reduction reagent, 2ml

FP-CG4891 1 kit

Technical tip – ANTS fluorescence assisted analysis of oligosaccharides of human transferin

*Reference: The effects of ethanol on the glycosylation of human transferrin.Flahaut C, Michalski Jc, Danel T, Humbert MH and Klein A, Glycobiology 2003, 13, 191-198 Article

Oligosacharides relases by PNGase F from transferin were purified by affinity from normal or severe alcohol abuse patients, then labeled with ANTS. ANTS-labeled oligosacharides were analysed by PAGE analysis assisted by fluorescent detection with ANTS [Jackson 1994]. After elution from the gel, oligosaccharides were also characterized by MS.

Proteomics - BiochemistryProtein analysis

Carbohydrate Analysis/Detection kit

Product Cat# Qty

APTS (λabs/em.: 424/505nm; EC=7200M-1cm-1)2-aminopyrene-1.3.6- TriSulfonic acid Na salt, MW: 523.4

FP-33972A 10mg

ANTS (λabs/em.: 353/520nm)8-AminoPhtalene-1.3.6-trisulfonic acid, MW: 427.3

FP-46574A 500mg

1.5 EDANS (λabs/em.: 335/493nm) FP-46479A 1g2-AminoAcridone (λabs/em.: 354/442nm)used to modify carbonyl-containing biomolecules, in particular, for labeling carbohydrates and glycoproteins

FP-485135 25mg

Inquire for all our FluoProbes labels derivatized by hydrazide (reactive/carbohydrate). Over 60 fluorescent labels are available to cover luorescence requirements (spaning from 390nm to 800nm, up 500ns liftime,...)

Other fluorescent probes for glycoside analysis

Electrofluorogram B : healthy subjects. Arrows indicate a ladder of maltooligosaccharides (Gn where n indicates the number of residues).

Protein analysis




Colorimetric & fluorimetric Protein assays

Protein assays are required in any proteomics study to monitor protein preparation and to quantitate proteins before analysis (electrophoresis, MS, µarray...). Interchim provides robust colorimetric and fluorimetric assays to allow you gain time and results accuracy: BC Assay and LavaPep Assay are recommended for analyzing post-traductional modified proteins (see the tech tip).

Technical tip – Moving from Bradford, Biuret and Lowry methods to BC Assay and LavaPep assays for proteomics studies

People are often not aware of limitations using the so-called and popular Bradford method fo assaying proteins: because of high protein to protein variations of signal and poor linearity, results are often biased and not reproducible enough for fine proteomics analysis. This is due to the interaction type of the included dye, Coomassie, that binds preferentially to certain amino-acids, hydrophobic pockets... This becomes critical with post-transductionally modified proteins, protein mixtures, and digested proteins before MS analysis.

To that point, Uptima recommends to choose preferably BC Assay that as several advantages (see table below) and is general more versatile in most cases. It uses a reaction with peptidic bonds of proteins similar to Biuret and Lowry reagents (making it acceptable when previous works referred to Biuret or Lowry), but improved by the use of bicinchoninic acid. As a result, BC Assay is easier to perform, more accurate (better linearity/broader dynamic range), and more reliable to quantitate diverse proteins. The protein detection sensitivity of the BC Assay is slightly lower in the standard version, but has same or even better sensitivity than Bradford assays with the enhanced protocol, and with the microBC Assay version. In limited cases, the Coo Assay remained irreplaceable (quick procedure required, compatibility with reducing agents and some chelators without removing these substances (desalting, TCA precipitation)). Now Lavap method appears :

Amongst fluorescent protein assays to achieve higher sensitivity, we recommend the LavaPep assay : it provides ultimate sensitivity, higher dynamic range, compatibility with many substance usually interfering, and is ideal for proteolytic-digested peptides with MS analysis requirements.


Selection guide

Proteins analysis

Product Cat.# Suits especially to applications where are needed Limitations

BC Assay UP40840A - Accuracy &versatility- Compatibility, notably with detergents, bases, nucleic acids, lipids…- Large dynamic range (20-1500µg/ml)- Recommended to replace Biuret, Lowry methods, and even Bradford method (see technical tip)

Presence of reducing agents, and some chelators*

MicroBC Assay@562nm(540-590nm)

UP75860A - As BC Assay, plus - Higher sensitivity (0.5-500µg/ml)

Same as BC Assay

Coo Assay UPF86400 - Quick of use (<5min)- Compatibility with reducing agents- Sensitivity (0.5-20µg/ml) OR flexibility (up 2mg/ml)

Presence of many detergents, lipids, alkalis...Protein to protein variationsLow linearity

LavaPep Assay CH4191 - Ultimate sensitivity - Large dynamic range (0.04/0.1 - 160µg/ml)- Ease of use (60min)- Compatibility, notably with detergents AND reducing agents, nucleic acids,...- Peptides assays (before MS analysis)- Safe and environment free- Cost/performance-effective

Need a fluorimeter

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BC Assay kit - colorimetric protein assay

BC Assay is the premium quality and state of art improvement version of Biuret and Lowry assays. It has become the standard colorimetric in most labs, thanks its ease of use (one step), its unrivalled linearity and working range (0.5 to 2mg/ml), its compatibility with many agents starting with detergents (e.g. SDS) and nucleic acids, and its sensitivity (20µg/ml and down 0.5pg/ml with the microBC assay). It works fine including for glycoproteins (see page 49 tech tip). Detection occurs at 540-590nm (opt.@562nm).A modified Bradford assay (Coo Assay) and a fluorescent assay (LavaPep, see next page) are available as well.

BC Assay Cat# Qty

for 500/5000 (tube/microplate) determinations UP40840A 1 kit*for 125/1250 (tube/microplate) determinations UP40840B 1 kit* Contains 1L of reagent A, 25ml of reagent B, 10x1ml of BSA standard 2mg/ml

MicroBC Assay Cat# Qty

for 500/3400 (tube/microplate) determinations UP75860A 1 kit*for 50/340 (tube/microplate) determinations UP75860C 1 kit* Contains 250ml of reagent A, 250ml of reagent B, 12ml of reagent C, 10 x 1ml of BSA standard 2mg/ml

Coo Assay Cat# Qty

for 500/4000 (tube/microplate) determinations UPF86400 1 kit*for 125/1000 (tube/microplate) determinations UPF86401 1 kit* Contains 1L of reagent, 10x1ml of BSA standard 2mg/ml

Technical tip – Fluorescent Protein Detection

LavaPep, LavaPurple, and LavaCell use Epicocconone, a natural fluorescent compound from the marine fungus epicoccumnigrum. It yields a fluorescence shift from weak green fluorescence (530nm) to intense red/orange fluorescence (610nm) upon binding to proteins.

Fluorescence can be read by many platforms, such as fluorescence imager, fluorimeter, fluorescence plat readers, and laser scanner.

Hence, Epicocconone is great for demanding protein analysis (sensitivity, robustness, rapidness, low protein-protein variations...) in solutions (<40ng/ml, DTT AND detergent compatible: see LavaPep next page), electrophoresis gels and blottting (SDS-PAGE, IEF, 2D, WB: see LavaPurple p.80), and in cells (inquire for LavaCell stain).

. abs/em. spectra : blue: absorption, green : emission (free), red : emission(protein bound)

. Optimum excitation wavelengths : 405, 500 nm. Suitable light sources include green (e.g. 543, 532 nm) blue (e.g. 488 nm); violet (e.g. 405 nm) or UVA.

. Emission wavelength : The maximum emission is at 610 nm, irrespective of the excitation source.





For the accurate and sensitive determination of peptides, as well proteins in solutions

Performant :• can be used with excitation at 405-500nm, and emission at 560-610nm• is sensitive, down <100 ng/mL (peptide) and 40ng/ml (proteins).• has a linear wide dynamic range between 100 ng/mL and 160 μg/mL (over 3-orders

of magnitude)• accommodates a wide range of convenient assay volumes: 100 µL - 3 mL • has low peptide-peptide or protein-protein variability, improving accuracy for complex

samples

Convenient :• is easy to use: simply mix 1 part working solution with 1 part of sample. • is quick - data can be read within 60 mins • is safe, biodegradable, environmentally friendly, simple to dispose of *• requires no heating steps or time-consuming heating and reduction steps..• signal is not affected by light or temperature and remains stable for up to 6 hours • suits a wide range of fluorescence measuring instruments • is robust to many interfering compounds such as DNA, solubilisation reagents and redu-

cers

Flexible to most applications :• is suitable for measuring peptides from proteolytic digestions and most pure peptides• is compatible to post-traductionally modified proteins/phospho- & glyco-peptides• accommodates samples for 1-D and 2-DGE • does not modify precipitate or denature peptides that can be used in subsequent

assays• is compatible with downstream analyses such as MS and HPLC.• is amenable to N-term sequencing and to functional assays.• is more robust than other peptide assays and more cost effective than AAA.• is ideal for high throughput analysis

Technical tip – LavaPep is great for post-traductionnally modified proteins/phospho- & glyco-peptides

LavaPep assay outperforms colorimetric Bradford and Lowry assays for sensitivity and linearity for both glycoproteins and non-glyco-sylated proteins. LavaPep is 4-50 times more sensitive depending on the glycoprotein, due its special binding scheme that is not affected by the degree of glycosylation (probably thanks to the dyes hydrophilic nature and low molecular weight), combined to its intrinsec higher sensitivity.

LavaPepTM peptide & protein assays kit

Product Cat# Qty

LavaPep peptide&protein Assay CH4191 1 kit up to 2000 assays Contains : Reagent A (stain), Reagent B (buffer), for assay of proteins and peptides in solution

Kinetic of trypsin digestion

Related product

Product Cat# Qty

LavaDigestTM Protease Monitoring kit CH6251 1 kit up to 2000 assays offers a simple real time monitoring of protein digestions before analysis of formed peptides. It is suitable for all proteases, do not need derivatization (simplifying MS analysis), works for phospho- and glyco-proteins, perform in solution in real-time (allows kinetic studies, and to check complete digestion occured for accurate MS analysis), has a simple staining procedure, is compatible for mass spectrometry and can be used on laser or CCD based imagers.

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Phosphate Assays

Because Phosphate plays an very important role in biomolecules activation, and in biochemistry, assaying phosphates is useful to study Phosphatases, ATPases and several other enzymes, as well for ecological studies (water). Interchim provides 2 assay kits, based on a colorimetric and on a fluorescent methods.

Technical tip – Phosphate in the life Sciences

Phosphorus is found in biological systems, as a free phosphate ion in solution that is called inorganic phosphate (Pi =PO4

3−, MW: 94.97), and pyrophosphate (Ppi = P2O7

4-) to distin-guish it from phosphates bound in various esters such as adenosine phosphates and phosphorylated proteins or lipids.

However, phosphates are most commonly found in the form of adenosine phosphates, (AMP, ADP and ATP), of other nucleoside di- and tri-phosphates, and in DNA, RNA and proteins. Pi can be released by the hydrolysis of nucleoside di- and tri-phos-phates (so-called "phosphagens" in muscle tissue), releasing high amounts of energy of the phosphoanhydride bonds to activate biomolecules (phosphorylation of proteins) or catalyse biochemical reactions.Because of its important role in biological systems, phosphate availability may govern the rate of growth of organisms and is thus an important ecological parameter in the study of bio-resources (carencies, booms) and of pollution (water quality because of phos-phates content of detergents and fertilizers, eutrophication).

Colorimetric Phosphatase Assay

The phosphate is dosed by a colorimetric assay based on molybdate and malachite green dye. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600 - 660 nm) or on a plate reader. The assay provides an alternative to hazardous radioactive methods and other less sensitive colorimetric assays. Assays can be per-formed in cuvettes or conveniently in 96-well plates for high-throughput screening.The kit can also be used to estimate the phosphate content of proteins (phosphoserine or phosphothreonine post-translational modifications, after alkaline hydrolysis.

Product Cat# Qty

Phosphate Assay, MG methodOriginal method

IS2790 1 kit (ca 600 assays)

Phosphate Assay, MG Plus methodImproved end-point stable signal (not prone to precipitation)

CI4211 1 kit (1000 assays)

An assay for blots is also available (R09430)

Fluorimetric Phosphatase Assay

The assay use a new red fluorescent phosphate sensor, to measure the activity of any Pi-generating enzyme or Pi-absorbing process. Sensitivity (down 0.1µM) overcomes colorime-tric assays and equals radioactivity methods without their hazards. It is ideal for kinetics of phosphatase such as GTPases and ATPases. The assay can be performed in down 30min procedure either in absorbance (570nm) or fluorescence (Exc./Em.:540/590nm) in 96- and 384-wells microplate format, automated with no separation steps.

Product Cat# Qty

PhosphoWorksTM Fluorimetric Phosphate Assay Kit, *Red FluorescenceContains phosphate sensor (1 vial), Assay buffer (5ml), and KH2PO4 (1ml) CI4161 100 tests

A version (#CI4191) coupled to MESG method is available, providing UV absorption detection, 2µM sensitivity.

Fluorimetric ADP Assay

The assay use a new red fluorescent phosphate sensor, to measure the activity of any ADP-generating enzyme or ADP-absorbing process. Sensitivity (<0.2µM) overcomes assays based on monitoring of phosphopeptide formation or ATP deletion (i.e. Luciferase based), which have limitation including respectively time/efforts consumption to optimize peptide substrates or antibodies, and various interferences in assay. It has a broad ATP tolerance (1-300µM) that makes it ideal for kinase assays (screening, identifying and characterizing (kinetics)).The assay can be performed in down 30min procedure either in absorbance (570nm) or fluorescence (Exc./Em.:540/590nm) in 96- and 384-wells microplate format, automated with no separation steps.Product Cat# Qty

PhosphoWorksTM Fluorimetric ADP Assay Kit, *Red FluorescenceContains ADP sensor (1ml), Sensor buffer (2ml), ADP Assay Buffer (5ml), qnd ADP standard

C14161 100 tests

ATP assays are as well available (#S2841A) Phophoprotein stain kit (#CD6040) presented p.70.





Innovative electrophoresis : NEXT gels

The newest high-resolution PAGE System for Protein Separation

New Electrophoresis X’PRESS Technology (NEXT GEL™)

New Electrophoresis X’PRESS Technology (NEXT GEL™) PAGE provides superior protein separations, with full range resolution in a wide range of proteins size. It is cost-effective alternative to gradient gels, but it is easier to use and cheaper. Ready to use premixed solutions are stable for 6 months and can be kept handy on your bench top at room temperature. Just add TEMED and APS, mix, pour, insert comb and run gel.NEXT GEL™ is fully compatible with standard SDS-PAGE applications such as 1D & 2D gels, Western Blot Transfer, Protein Sequencing, MALDI Analysis, and common stain methods.

Benefits

Ultra-Fine Resolution : • Resolve 14.4 kDa from 14.2 kDa using a 10% NEXT GelTM mini-gel.

Save Money : • NEXT Gels are cheaper and easier to make than gradient gels.

Saves Time / Ease-of-Use : • Stable for months • No stacking gel required. • Requires less than a minute to pour a gel.

To order NEXT Gel™ ready-to-use acrylamide solutions and buffer Description Fine resolution range Cat.#/100ml Cat.#/500ml (1)

NEXT GelTM 5% 50-200 KDa GS4270 GS4271 NEXT GelTM 7.5% 20-100 KDa GS4280 GS4281 NEXT GelTM 10% 10-70 KDa BG6290 BG6291 NEXT GelTM 12.5% 10-50 KDa GS4290 GS4291 NEXT GelTM 15% 5-40 KDa GS4300 GS4301 NEXT GelTM Running Buffer, 20X GS4310 GS4311LP-NEXT Gel™ 14 – 3 000 KDa BI6150 / 1kit (3)

(1) Each package of 500ml contains reagents for 50 mini and 20 regular gels.(2) This ready to dilute buffer will achieve high resolution on a wide range of protein fragment sizes. (3) This Kit includes : NEXT Gel™ sample buffer 4X ,NEXT Gel™ running buffer, 20X Agarose High Resolution Protein (HRP)

Large Protein NEXT GEL™ (LP-NEXT™)

Large Protein NEXT GELTM (LP-NEXTTM) electrophoresis is a novel and UNIQUE system to analyze large molecular weight proteins via horizontal agarose gel electrophoresis.

• Specifically formulated for separating large MW proteins under denaturing conditions • Resolve proteins from 14 000 up to 3 millions Daltons on a single gel • Superior protein resolution compared with most commercially available gels·

Uses readily available horizontal mini submarine agarose gel equipment • Compatible with all supporting technology such as western blotting, N-terminal sequen-

cing, MS MALDI, etc • Saves time as easy to run as a DNA agarose gel • Contains sufficient reagents to perform 35-50 mini gels • Stable at room temperature for up to 1 year

Find more electrophoresis reagents in our BioScience catalogue pages B194-B203, including High quality and economic acrylamide solutions, biochemicals, ready to use buffers...

Intermolecular Cross-linking of Myosin Heavy Chain by Glutaraldehyde.Samples analyzed with LP-NEXT GEL™ 1% Agarose HRP followed by silver staining.Lane 1 : Myosin (minor bands are aggregates).Lane 2-4 : Myosin cross-linked with Glutaraldehyde for various time intervals.

Soon available in pre-cast gels.

10% Next GelTM 12 % Competitor

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GeBa Electrophoresis system / pre-cast gels

An economic, easy-to-use horizontal electrophoresis system with resolutive and cost-ef-fective pre-cast gels

• User-friendly protein electrophoresis system. • Easy-to-use: horizontal apparatus (deposit sample with standard pipette tips). No special loading tips needed, agarose standard-loading tips used. No leakage of running buffer from inner tank to the outer tank. • Running buffer saving, use 150 ml only, with the same standard Tris-glycine running

buffer even for peptides • High resolution: sharper bands provide clear accurate results. • Multiple applications: proteins, peptides * • Cost-effective & 12-months shelf-life guarantee.

*NEW : works also for nucleic acids !

The GeBa Electrophoresis system is a novel semi dry horizontal pre-cast gel system. Each research can have its own electrophoresis units instead of sharing costly systems! The hori-zontal design simplify considerably handling procedure :

No more cumbersome assembly of the gel with the running apparatus. No adaptors required Robust sample wells dividers eliminate damage when removing the comb and loading, and don’t deform or fall

over. Effortless cassette opening compared to competitor pre-cast gels.

The GeBa Electrophoresis system is a very versatile system, for protein, peptide as well nucleic acids ! Gels can be removed easily, stained, electroeluted for MS, or electrotransfered using standard membrane transfer buffer.

Comparison of GeBaGel system with standard pre-cast gel (vertical) systems

Product Comb color Cat.# Unit

10% Protein GeBaGel /Proteins Blue BI9690 8 u12% Protein GeBaGel /Proteins Green BI9700 8 u4-12% Protein GeBaGel /Proteins Violet BI9710 8 u8-16% Protein GeBaGel /Proteins Red BI9720 8 u17% Protein GeBaGel /Peptides Black BI9730 8 uGEBARUNNER Electrophoresis system BI9740 1 uStarter kit : 1 GebaRunner + 8 gels BI9741 1 kit

Competitor electrophoresis systems GeBaGel system

Mode Vertical HorizontalSystem assembly Assembly is awkward and need skill Plus and playSample loading Nedd skill, special loading tips EasyLeakage possibility Exists from time to time Not existsVolume of buffer / run ProteinPeptide

500 mlcostly buffers (MOPS, MES, HEPES)costly Trictine or MES buffer

only 150 mlregular Tris. Glycine bufferregular Tris. Glycine buffer

Home made gels Pre-cast gels pH8.7 Pre-cast gels pH7 GeBaGels

Storage --- - ++ ++Resolution +++ ++ - +++Cost +++ + -- ++Handling --- + + ++Applications ProteinsPeptides Proteins Proteins Proteins, Peptides,

DNA, RNA




Proteomics - BiochemistryProteins analysis

ProSaveTM Protein Gel Stain

Our ultra-fast, easy, sensitive and flexible gel stain : Stain the gel, and save the proteins !

Main features and benefits

• Quickest staining : 5-10 min • Highest sensitivity : nanograms of protein, and DNA • Good dynamic range • Cost effective (cheaper than fluorescent methods) • Non toxic, and stable reagent. • Do no modify or interact with proteins, to keep them in better state (non-denaturing

gels) • Detect all proteins (including glyco-, phospho-, and lipo-proteins and other proteins

difficult to stain), with minimal protein to protein variations • Fully and quickly reversible: 5-7 min • Fully compatible with more downstream applications : . Re-staining by any other method . Electrotransfert onto PVDF or NC membranes for immunoblotting . Electroelution for protein recovery . Mass Spectrometry analysis (more spots detected) . Amino acid analysis . N-terminal sequencing

ProSave is definitively the best solution to fulfill your requirements of protein staining in acryla-mide gels ! Not like conventional methods, such as silver stain, Coomassie stain as well a Ruby stain, taking a few hours to complete. ProSave enables you to get high quality results within just 5 min, without overstaining on speckles.ProSave can detect as little as 1ng in SDS-PAGE gels, that is much more sensitive than Coo-massie stains (CBR); it compares to Silver stain and Fluorescent Rubis stain. Furthermore, the dynamic range is good, all proteins are stained (no preference as exist for glycoprotein by silver, or lower staining of "difficult proteins" with fluorescent stains).ProSave is a robust method for proteomics 2-D gels staining, and yields usually higher rate of good annotation. As no component bind to proteins, it is fully and quickly reversible, and ideal for mass spectrometry.

Product Cat# Qty

ProSaveTM Protein Gel stain1 kit provides sufficient quantity of 2 solutions (a sensitizer and a developping) to stain 20-25 minigels.

BP7121 1 kit (ask for trial offer)

Easy and quick use

Excellent sensitivity

ProSave stain Silver Stain

Higher rate of good annotations in MS

MALDI-TOF spectrum of rabbit phosphorylase b spot (cut, prepared for MS, ionized fragments were annoted)

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LavaPurpleTM protein gel & blot stain

The most versatile and sensitive protein stain for gels AND blots, with superior results for proteomics requirements

• Ultimate sensitivity : detect as low as 50pg • Low protein to protein variability • High signal to noise

• Safer to use & simpler to dispose of (biodegradable - not heavy metals) * • Simple and quick : 1-3 Hr (gels) or < 45min (blots) • Suited to automated high throughput systems

LavaPurple : 1076 spots Rubys stain: 877 spotsRat microsomal proteins focused in 17 cm pH 3 – 10 IPG strips and separated in large format 2D gels. Replicate gels were stai-ned using LavaPurple and SYPRO Ruby. The spot number in each gel was calculated by the Malanie 3 software (Gene Bio). (Taken from Mackintosh et al., 2003, Proteomics 3, 2273 – 2288.)

LavaPurple Rubys stainLow molecular weight markers two-fold diluted from approxi-mately 128–1 ng blotted to Hybond-P (PVDF) and stained with LavaPurple Total Protein Stain. The minimal level of detection was at least 1 ng of the protein concentrations (as loaded into the gel ). Staining of the gel after protein blotting revealed that not all protein had transferred to the membrane.

Optimum excitation & Emission wavelengths : 405, 500 nm / maximum of 610 nm.

Suitable light sources : include green (e.g. 543, 532 nm); blue (e.g. 488 nm); violet (e.g. 405 nm) or UVA.

Suitable filters : include the 610 nm band pass or 560 long pass.

Product Cat# Qty

LavaPurpleTM protein gel & Blot Stain* 5ml dilutes to 1L, allowing staining of ca 20 minigels or 2-3 large format gels

674331674332674333

5ml 200X*25ml 200X*100ml 200X*

Blot staining applications

• 16-Fold more sensitive than Rubys stains

• Works fo all blots (WesternB., 2D, IEF)

• Low background and no speckling • Compatible with MS** and Edman-

based sequencing • Compatible with functional analysis

and antibody staining

Gel staining applications

• Linear quantitation over 4 orders of magnitude

• More compatible with sequencing, functional analysis, and mass spec-trometry **

• More real protein spots / less false positives on 2D gels

• Staining can be reversed easily • Multiplex compatible with DIGE

(Cy™), Phosphoprotein (PQ), silver and Coomassie staining

Product Cat# Qty

LavaPurpleTM protein gel & Blot Stain Kit* Contains : Reagent stain (200 X), and reagents buffer, for staining proteins and peptides in electrophoresis gels

67433A67433B67433C

1 small kit (20 minigels)1 medium kit (100 minigels)1 large kit (400 minigels)

Highlight – LavaPurple is great for post-traductionally modified proteins/phospho- & glyco-peptides

LavaPurple assay outperforms Ruby and Krypton stains for sensitivity and linearity with both glycoproteins and non-glycosylated proteins. LavaPep is 8 and 32 times more sen-sitive depending on the glycoprotein, due to its special binding scheme that is not affected by the degree of glycosylation (probably thanks to the dye hydrophilic nature and low molecular weight), combined to its intrinsec higher sen-sitivity). Rare proteins lost with other stains, can be found! Also, staining is not inhibited by the degree of glycosylation compared to Ruby stain, even highly glycated proteins are detected. Finally a greater number of peptides are detected by analysis of gels and by MS analysis of digested proteins eluted from of 1D and 2D electrophoresis gels.

* No heavy metals unlike Rubis and silver stains

** More compatible than competitor products: see figure below

Phosphoprotein in gel stain kit : see page 70 product #CD6040





Blotting glycated proteins with Protran®

Protran® blotting membranes are not only the world-wide reference for Western-Blotting, also specially excellent for glyco-proteins and protein interactions studies.

Glycosylation is well-known to modify proteins in a proper three-dimensional structure allowing to address them to the right location in cells. Glycosides are also involves in many protein recognition sites of enzymes, receptors...

One approach for discovering new carbohydrate-recognizing proteins in the proteome, and for mapping carbohydrate recognition structures in the glycome, are arrays of oligosaccharides [1]. The figure opposite shows the highly sensitive detection of carbohydrates on Protran membranes. A quantitative immunostaining experiment clearly indicates a more than 10-fold higher signal intensity for oligosaccharides (as NGLs*) on Protran nitrocellulose.

[1] The NGL technology generates lipid-linked oligosaccharide probes from glycoproteins and polysaccharides which are parti-cularly suitable for the arraying of oligosaccharides. Fukui S., Feizi T., Galustian C., Lawson A. M. and W. Chai. Oligosaccharide microarrays for high-throughput detection and specificity assignments of carbohydrate-protein interactions. Nature Biotechnology 2002, Vol. 20, 1011-1017

Protran® offers also highest sensitivity for protein-protein interaction studies. Using fluores-cence labeled probes and protein samples spotted on Protran enables analysis of signals from protein-protein or protein-DNA interactions with high sensitivity of detection (ask for application notes).

Protran suits Western-Blotting and dot/slot blots, as well as nucleic acids (N.A.) in Southern- and Northern-blotting, hybridization fo colonies and plate lifts. Protran is superior to PVDF and Nylon for proteins and carbohydrates, in particular for peroxidase/ECL based detections.Its purity is unrivalled (no cellulose acetate, that reduce protein binding). Its mechanical resistance facilitates handling (avoids stretches). At the opposite of PVDF, no pre-wetting in methanol is required. The retention of biomolecules during transfer is excellent (3 porosity levels depending of molecular size).

Immuno detection of oligosaccharides immobi-lized as NGLs [1] on Protran® BA85 (Data are kindly provided by Professor Ten Feizi, Imperial College London, Northwick Park & St Mark’s Hospital Campus Harrow, Middlesex, UK.The method is described in [1].)

Protran Nitrocellulose membranes

Nucl. ProteinsAcid >20 kD <20 kD <7kD

Sheets of 20 x 20 cm

Roll of 3 m x 30 cm

BA85 NC 0.45 µm ++ ++ + - BN3802, 25uBN3801, 5u

U60640, 1u

BA82 NC 0.2 µm + + ++ - S31442, 25 u BN3841, 1uBA79 NC 0.1 µm - - - ++ BN3652, 25 u BN3831, 1u

Ask for other forms (disks, sheets), for nylon (Nytran®) or PVDF (TotalBlot™) membra-nes.

Nitrocellulose membranes, 0.45 µm pore size, from different manufacturers/suppliers were cut into pieces of 15 x 100 mm. Tensile strength was measured (according to DIN 53 112, part 1) in an automatic tension and pressure recording apparatus. Data indicated are the mean of at least 4 independent measurements.Membrane thickness : A = 130 µm, B = 130 µm, C = 160 µm, D (mixed ester) = 160 µm and Protran BA85 = 125 µm.

Superior tensil strength

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One-Step Western Blot Kit

Shortens the time of Western Blot analysis >4 times • rapid and simple procedure (one step) • no saturation nor secondary antibody • sensitivity and reproducibility comparable to classic WesternBlot

Simply incubate the blot with your primary antibody in the WB solution, your blot is ready for development with ECL UptiLightOne or any other HRP substrate.

Product Cat# Qty

Step Western kit (Mouse) BU9490 5 to 25 runsStep Western kit (Rabbit) BU9500 5 to 25 runs

ReliaBLOTTM IP/Western Blot

Improves the WB detection of immunoprecipited proteins.

• Efficient : Erase unspecific WB detection due to heavy chains of Igs = cleaner results in WB = Accurate detection of usually masked bands. • Sensitive : Low levels of proteins are detected. • Simple : Do not need to change your IP protocol.

ReliaBlot is compatible with Protein A and G, and avoid time-consuming cross-linking of Ab. It is optimized for Rabbit Abs in Western Blot after IP (mouse and goat reagents coming soon).

Cat# Qty

ReliablotTM IP/Western Blot Reagents BY1150 20 testsBY115T 5 tests*

See alternative method with IPEX ImmunoPrecipitation kit page 48





UptiLightTM, the best of ECL detections

Uptima is pleased to present UptiLight™ substrates for Enzyme ChemiLuminescence (ECL). UptiLight is an exceptional formulation of luminol, the best chemiluminescent substrate for peroxidase. Uptilight produces in presence of HRP a particularly intense luminous emission thanks to different proprietary agents increasing the photonic emission yield.

UptiLight blotting substrate is optimized for the detection of the immobilized peroxidase, especially in Dot, Western, Southern and Northern- blotting techniques on nitrocellulose, nylon and PVDF. The signal is registered on radiographic films, by scanners or with a CDD camera. Excellent results are obtained with shorter exposure duration than competitor products. Op-timal sensitivity/background/time of operating is usually achieved in only 1-15 min. The good stability of signal allows the multiple re-exposure of the radiographic film in order to refine the intensity of revelation, or in order to obtain several copies.

UptiLight comes in following convenient formats :

• UptiLight classic kit #UP99619 is the most cost effective ECL product in the market. • UptiLightOne spray #BM4961 is a breakthrough put in the labs : just spray this ready-

to-use one component reagent ! This format is dedicated to maximal convenience and gain of time.

• UptiLight HighSensitive kit #98490 is a very sensitive and economic ECL substrate for detections in the pico to mid-femto gram range detections.

• UptiLight UltraSensitive kit #58372 provides ultimate sensitivity for most demanding applications, covering the femto gram range detections.

UptiLight ELISA substrate is designed for assays in microplates, especially for high through put screening assays in proteomics, immunochemistry and cell biology. It comes in 2 sensitivity levels:

• UptiLight ELISA HighSensitivity kit #36349 • UptiLight ELISA UltraSensitivity kit #99620

UptiLight Elisa HighSensitivity Qty

36349A 200 ml36349B 600 ml36349C 2 000 ml

UptiLight Elisa Ultra Sensitivity Qty

996201 60 ml996202 120 ml996203 300 ml

UptiLight Classic Qty

UP99619A 500 ml

UptiLight WB HighSensitivity Qty

98490A 200 ml98490B 600 ml98490C 2 000 ml

UptiLight WB UltraSensitivity Qty

58372A 60 ml58372B 120 ml58372C 300 ml

UtpiLightTM solutions at a glance

UptiLight™ ELISA ECL Substrates UptiLight™ Blotting ECL SubstratesApplications :microplates assays using HRP (ELISAs)

Applications : Western-, Southern-, Northern-Blotting with peroxidase (HRP)

UptiLight One (1 component)

BM4961 2 500 cm2

BM4963 Dropper 5 000 cm2

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Proteomics -BiochemistryProtein analysis

Superior contrasted images

Superior conveniencySuperior sensitivity

Highlights

UptiLightOne (1 component, spray) :

Competitor ECL(2 components, mixed) :

Cost-effectiveness

• Save your budget, and gain sensitivity, choosing the UptiLight™, the brilliant solution !

Superior reagent stability

UptiLight E (Competitor)

E+ (Competitor) SP (Competitor)

UptiLight HS #99620 (2 components, mixed)





High quality Biochemicals

More detergents page B2-B10 of our BioScien-ces catalogue

Product Qty Biotech grade Proteomics grade

SDS powder 100 g 08938A GS3750250 g 08938B GS3751500 g UP08938C GS3752

SDS, 20% (w/v) solution 200 ml 896820 89682A 500 ml UP896826 89682B

Tween 20, Oxidant Free, 10% solution 5 x 10 ml UP158740Brij-35 1 Kg 09187A 09187KBrij35, Oxidant Free, 10% solution 5 x 10 ml UP091870Triton X100 1 L 15851B 521128Triton X100, Oxidant Free, 10% solution 5 x 10 ml UP521121Triton X114 1 L 15852F 15852DTriton X100, Oxidant Free, 10% solution 5 x 10 ml UP158528n-Octyl-β-D-glucopyranoside, UltraPure 1 g UP263700 26370B n-Octyl-β-D-thioglucopyranoside, UltraPure 1 g UP602080 60208BHecameg 5 g UP785480Thiourea 100 g 03004HUrea 500 g UP0311903 03190EUrea 8M solution 250 ml N13830 N13831Guanidine HCl UP018380, 500 g 01838L, 250gGuanidine 8M solution 4 L GS3780

Protease Inhibitors (more page A386 of our BioScience catalogue)


AEBSF UP¨401071, 1g GS4071, 250mgAntipain diHCl 5 mg UP257317 25731CAprotinin 10 mg UP185582 18558DBenzamide HCl 25 g 003051 003059Bestatin 10 mg UP300991 300995E-64 5 mg UP789581 78958EEDTA Na salt 500 g T32141Leupeptin 5 mg UP827721 827728Phosphoramidon UP348115, 5mg 348118, 5gPMFS 25 g UP147374 GS3920Protease inhibotor coktail – general use 1 ml 374723Protease inhibotor coktail – general use 1 ml 374723Protease inhibotor coktail – general use, with EDTA 1 ml 374724Protease inhibotor coktail – Mammalian use 1 ml AN0990Trypsin inhibitor (soybean) 1 g N15150 N15152α-Chymotrypsin 1 g 571765

Buffers (more page A386 of our BioScience catalogue)


BES 100 g T31611 BA7850Boric acid 1 Kg UP070440 10853BGlycerol 1 L 047620 04762KGlycine 1 Kg UP018225 01822SMOPS free acid 100 g UP06200 06200QMOPS Na salt N13431 N13433nTris base UP031657, 1kg 03165P, 500gTris HCl UP09154E, 1Kg 09154P, 500gTBS with Non-Fat Powdered Milk 3% (42 g/1 L) 5 pk GS4160 TBS with BSA 1% (22 g/1 L) 5 pk GS4170 TBS with Tween® 0.05% (12.5 g/1 L) 5 pk GS4200 PBS with Non-Fat Powdered Milk 3% (39.8 g/1 L) 5 pk GS4180PBS with BSA 1% (19.8 g/1 L) 5 pk GS4190 PBS with Tween® 0.05% (10.4 g/1 L) 5 pk GS4250

DetergentsSDS (MW : 288.38), an anionic detergent, is a critical reagent in many molecular biology ap-plications. For example, contaminating levels of C16-alkyl sulfate particularly affect protein renaturation, and contaminating UV absorbing materials affect detection sensitivity. Additio-nally, heavy metals/ chloride contaminants affect separation and enzymatic activities.

Our Biotechnology Grade SDS is especially high in both purity and C12 content., to suit the most demanding biotech applications., molecular biology and genomics.

Biotech grade :Excellent batch to batch reproducibilityPurity (HPLC) > 99%Content in C12 : > 99%Nuclease, RNase and protease freeOD260 and OD280 (3% solution in water) < 0.1Chlorides < 0.1%Copper, Lead < 5ppm

Protease coktails contains a mix of AEBSF, Aprotinin, E-64, Bestatin and Leupeptin (+Pepstatin for the Mammalian use item)

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Trademarks and legals

BC Assay™ ..................................... InterchimControlledAmine™ ......................... InterchimCoo Assay™ ................................... InterchimCy™ ................................................. AmershamHecameg® ........................................ VegatecIMMUNOcatcher™ ......................... CytosignalKinome™ ......................................... AbgentLavaPep™ ....................................... FluorotechnicsLavaPurple™ ................................... FluorotechnicsLP-NEXT™ ...................................... AmrescoNEXT GEL™ ................................... AmrescopLIVE™ ........................................... MirusProSave™ ....................................... InterchimProtran® ........................................... Schleicher&Schull (Whatman)ReliaBlot™....................................... (patent pending)siXpress® ......................................... Mirus Inc (Takara)TransIT® ........................................... Mirus Inc (Takara)Tween® ............................................ ICI AmericaUptiLight™ ...................................... InterchimUptiTip™ .......................................... Interchim

Trademarks and legals




Conditions générales de vente1 - GénéralitésTous nos produits sont destinés à des usages professionnels, la vente aux particuliers est refusée.Les conditions générales de vente sont automatiquement communiquées à tout nouveau client à sa première commande. Sans contestation dans un délai de 7 jours calendaires, Interchim considérera que son client a pris connaissance des conditions générales de vente et qu’il les accepte pour la première commande et les suivantes.Sauf convention spéciale écrite, la passation d’une commande par le client implique son acceptation automatique et formelle aux présentes conditions générales de ventes. Aucune disposition contraire à nos conditions générales apportées sur les bons de commande, lettres, accusés de réception, conditions générales d’achat ou autres documents émanant du client ne saurait être opposée à Interchim si elle n’a pas été expressément acceptée par Interchim. Toute disposition qui déroge ou complète les présentes conditions générales de ventes sera considérée comme acceptée par le client en l’absence de toute contestation écrite de ce dernier.

2 - Commandes Toute commande peut être passée par écrit, fax, e-mail ou téléphone (Hot-Line : Interchrom 04 70 03 73 09 Interbiotech 04 70 03 73 06 Interbulk 04 70 03 73 01). Toutefois les commandes téléphoniques ne sont acceptées que si un numéro de commande est donné par le client, dans ce cas la commande doit être confirmée par écrit. Interchim se réserve le droit de refuser une vente si la réglementation française, celle du pays d’achat et ou de vente n’est pas respectée par : le client, le destinataire final, les intermédiaires, les transporteurs.

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6 - RetoursInterchim a mis en place une procédure de retour. aussi aucun retour ne peut être effectué sans l’accord écrit d’Interchim. Une des conditions d’acceptation des retours est que le produit soit à l’état neuf et ré-emballé dans son emballage d’origine sans altération (étiquettes, écritures......).

Le client engage sa responsabilité civile et ou pénale si le produit retourné est altéré par des matières chimiques ou biochimiques ou biologiques ou toutes autres altérations de nature à constituer un risque pour les tiers et ou pour le personnel d’Interchim. Sauf erreur manifeste d’Interchim, les frais de port de retour sont à la charge du client. En cas d’erreur du client et après acceptation d’Interchim Les frais de stockages sont de 20 % du montant du produit avec un minimum de 45 €. (Ces frais de restockage incluent les frais de douane payés et non récupérables, les frais bancaires, les frais de retour aux fournisseurs et leurs propres frais de restockage).

7 - Clause de propriété Interchim se réserve expressément la propriété des produits livrés jusqu’au paiement intégral du prix de vente et des intérêts éventuels, frais, accessoires. A cet égard, ne constitue pas un paiement au sens de la présente disposition, la remise de traite, chèque ou tout autre titre créant une obligation de payer. Le paiement ne pourra être considéré effectué que lors de l’encaissement effectif du montant de la facture par Interchim. Le défaut de paiement de l’une des échéances pourra entraîner la revendication du matériel par Interchim. En cas de cession du matériel non payé, le client s ‘engage à la première demande d’Interchim, à lui transférer les créances qu’il détient sur les sous acquéreurs et ce à concurrence des sommes encore dues à Interchim.

8 - Clause résolutoireToute inexécution totale ou partielle par le client de l’une de ces obligations, le non respect d’une échéance quelconque de paiement pourra entraîner au gré d’Interchim d’une part la déchéance du terme et en conséquence, l’exigibilité immédiate des sommes encore dues à quelque titre que ce soit, ainsi que la suspension de toute livraison et d’autre part la résolution des contrats en cours.La résolution des contrats interviendra de plein droit et sans formalité judiciaire, à l’issue d’une période de huit jours calendaires après l’envoi au client d’une mise en demeure de payer par lettre recommandée avec accusé de réception, mentionnant l’intention d’utiliser la présente clause et restée sans effet, sans préjudice des autres droits d’Interchim. En cas de la mise en oeuvre de la présente clause résolutoire, Interchim ou son mandataire est expressément autorisé à pénétrer dans les locaux du client pour reprendre possession des produits concernés par l’article .

9 - Droit applicable et attribution de JuridictionLes présentes conditions de ventes ainsi que les contrats conclus en application sont régis par le droit français à l’exclusion de la convention de Vienne du 11 avril 1980 relative aux contrats de vente internationale de marchandises. Tout litige relatif à l’interprétation ou à l’exécution des conditions générales de vente et tout contrat conclu avec un client, qui ne pourrait dans un premier temps être résolu à l ‘amiable sera de la compétence exclusive du tribunal de commerce de Montluçon.

10 - Force majeureInterchim n’encoure aucune responsabilité en cas de force majeure. L’ exécution de l’obligation est retardée jusqu’à la cessation du cas de force majeure. Si la force majeure se poursuit au delà des deux mois, le contrat peut être résolu sans indemnité de part et d’ autre sur la demande de l’une des deux parties. La force majeure est un évènement imprévisible, insurmontable et extérieur à Interchim et faisant obstacle à son fonctionnement normal.

11 - Utilisations des ProduitsA l’exception des produits pharmaceutiques à usage pharmaceutique vendus à l’industrie pharmaceutique, tous nos produits sont uniquement destinés à la recherche et ne doivent en aucun cas être utilisés comme médicaments, cosmétiques, produits agricoles ou pesticides, additifs alimentaires, ou produits d’entretien. Ils doivent être utilisés par des personnes compétentes avec toutes les précautions habituelles, en accord avec les données, de la littérature

12 - Toxiques et substances dangereusesLes stabilités ou toxicités sont données d’après les informations de nos producteurs sous leur seule responsabilité, y compris en cas de caractéristiques ou de classes de toxicités erronées. L’absence d’une mise en garde ne doit pas être considérée comme une marque de sécurité car nous tenons à vous rappeler que pour bon nombre de substances chimiques, biologiques, biochimiques il n’existe pas de renseignements explicitant tous les dangers possibles. En conséquence, le client a la responsabilité de vérifier les dangerosités et d’effectuer les recherches nécessaires pour connaître les dangerosités induites par l’utilisation des produits achetés à Interchim. Les produits dangereux ne sont pas identifiés dans le catalogue. C’est de la compétence exclusive du client de contrôler la nature du risque attaché aux substances dangereuses. Le client a aussi le devoir de prévenir ses propres clients et les intermédiaires (transporteurs manutentionnaires) du risque induit en utilisant et ou manipulant les produits.

13 - Garantie La garantie est strictement limitée à la remise en état ou au remplacement des produits affectés d’un vice caché ou d’un défaut de conformité , à l’exclusion de tout autre dédommagement à quelque titre que cela soit. Le produit devra être remis à Interchim ou à un transporteur en parfait état dans son emballage d’origine, et le client devra se conformer aux instructions d’Interchim pour effectuer son retour, selon les modalités de l’article 6.

14 - Environnement Une ecotaxe sera appliquée suivant les directives relatives aux DEEE, conformément aux lois en vigueur pour les équipements électriques ou électroniques mis sur le marché à partir du 13 Août 2005. L’organisation, l’enlèvement et la destruction des produits demeurent à la charge de l’utilisateur final. Interchim reversant la contribution collectée aux organismes de recyclage agréés.

15 - Modifications des conditions générales de ventes Interchim se réserve le droit de modifier à tout moment les présentes conditions générales de ventes et en tel cas les conditions modifiées s’appliquent à toutes les commandes passées après la date de modification, même pour les commandes complémentaires ou connexes à des opérations antérieures.

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