New Levels of Accuracy for All Your Proteomics Research · 2016. 2. 28. · SEP010 Seppro IgY 14 Spin Columns 500 μL resin bed volume 15–20 μL (pooled normal human plasma or serum)

Bioprecision.Introducing new levels of accuracy for all your proteomics research.

biomapping

Protease Inhibitor Cocktails

Magnetic Beads

Seppro®

AQUA Peptides™

UPS

Panorama® Arrays

biomapping

Sigma Magnetic Beadsmean faster protein recovery and more accurate quantitation

The magnetic properties of Sigma’s agarose beads allow for:

• Very rapid separation• Significant accelerated manipulations, such as repetitive washings• Processing of multiple samples performed in plate formats

This leads to:

• Faster experimentation• Better reproducibility• More accurate quantitation of the proteins of interest

Overview of Magnetic BeadsThe magnetic properties allow for very rapid processing. These properties aid in manipulations, such as repetitive washings, and recovery of the protein bound beads. This leads to faster recovery, experimental reproducibility, and more accurate quantitation of the proteins of interest.

The matrix for these magnetic beads is 4% beaded agarose, with a diameter of 20–75 μm, with an average diameter of 50 μm. Superparamagnetic iron is impregnated within the bead. Beads are supplied in 50% glycerol with preservative and are stable for 2 years at recommended storage temperatures.

Sigma now offers magnetic beads for immunoprecipitation, protein purifications and the study of protein-protein interactions.

Magnetic Agarose BeadsFor Epitope Tag Protein Purifications

ANTI-FLAG® M2 Magnetic BeadsThe FLAG® Expression System is a proven method to express, purify and detect recombinant fusion proteins.

The ANTI-FLAG® M2 Magnetic Bead (M8823) is composed of murine derived, anti-FLAG M2 monoclonal antibody capable of binding to fusion proteins containing a FLAG peptide sequence at the N-terminus, Met-N-terminus, or C-terminus locations in mammalian, bacterial, and plant extracts. Binding attributes are similar to the standard ANTI-FLAG M2 Agarose (A2220).

More information regarding FLAG and FLAG magnetic beads can be found at sigma.com/flagmag

HIS-Select® Nickel Magnetic Agarose BeadsHIS-Select Nickel Magnetic Agarose Beads (H9914) are designed to capture proteins with histidine tags while exhibiting low non-specific binding of other proteins. They can be used in a wide variety of cell lysates and other biochemical solutions in a manner similar to the standard HIS-Select Nickel Affinity Gel (P6611). Both contain a proprietary, quadridentate chelate, which prevents non-specific binding and nickel leaching.

The capacity of the magnetic beads is typically greater than 10 mg/ml of packed gel, as determined with ~30 kDa his-tagged protein.

More information regarding HIS-Select and HIS-Select magnetic beads can be found at sigma.com/hismag

To learn about our complete recombinant protein expression workflow offerings, visit sigma.com/rpe

http://www.sigmaaldrich.com/life-science/proteomics/recombinant-protein-expression/purification-detection/flag-system.htmlhttp://www.sigmaaldrich.com/life-science/proteomics/recombinant-protein-expression/purification-detection/his-select/magnetic-agarose-beads.htmlhttp://sigma.com/rpe

Order 800-325-3010 Technical Service 800-325-5832 3

Broad Spectrum Protease InhibitionIn Solution or Tablet Formats

Protease Inhibitor CocktailsProtect the Integrity of Your Proteins

Four main classes of proteolytic enzymes have been routinely used to describe proteases:

• Serine Proteases• Cysteine Proteases• Aspartic Proteases (Acid Proteases)• MetalloproteasesAt the onset of tissue or cell lysis, the extract is teeming with proteolytic enzymes, which will degrade proteins. During isolation and characterization, one or all four classes of proteases may pose a

threat to the fate of a protein. It is equally important to have isolated proteins that are functional and compatible with subsequent downstream applications. Broad spectrum protease inhibitor cocktails have been developed to help protect the integrity of proteins.

Protease Inhibitor Cocktails and TabletsTablets are water soluble, contain no organic solvents (like DMSO), and protect against all four protease classes. Available in General Use and EDTA-Free formulations, each tablet reconstitutes to 100 mL, and is recommended for the inhibition of the proteases present in a 20 g cell mass.

P2714 S8820 I3911 P1860 P8465 P8340 I3786 P8849 S8830 P8215 P9599 MSSAFE

ApplicationGeneral

UseGeneral

UseGeneral

Use (ACF)Tissue

CultureBacterial

Cells

Mammalian Cells and Tissues

Mammalian Cells and

Tissues (ACF)Histidine-Tagged

ProteinsFungal/

Yeast CellsPlant Cells

Mammalian Cells and Tissues

Product Format

Lyophilized powder (water

soluble)

Tablet (water

soluble)


soluble)Solution in DMSO

Lyophilized powder (DMSO soluble) Solution in DMSO

Tablet (water

soluble) Solution in DMSO


soluble)

AEBSF ✓ ✓ ✓ — ✓ ✓ ✓ ✓ ✓ ✓ ✓ —

Aprotinin ✓ ✓ ✓ ✓ — ✓ ✓ — ✓ — — ✓

Bestatin ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ — ✓ ✓

EDTA ✓ ✓ ✓ — ✓ — — — — — — —

E-64 ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ —

Leupeptin ✓ ✓ ✓ ✓ — ✓ ✓ — ✓ — ✓ ✓

Pepstatin A — — — ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓

Phosphoramidon — — — — — — — ✓ ✓ — — ✓

1,10-Phenanthroline — — — — — — — — — ✓ ✓ —

Elastinal — — — — — — — — — — — ✓

Nafamostat mesylate — — — — — — — — — — — ✓

Antipain — — — — — — — — — — — ✓

biomapping

Seppro DepletionUnmask Low Abundance Proteins

Seppro®The Most Complete Human Protein Depletion System Available

• Depletion from a variety of sample types• Spin-Column and LC• Specific capture of target proteins• Complete solution, reagents, and techware includedThe Seppro depletion technology allows for the removal of several highly abundant proteins from a variety of biological samples. The use of Avian polyclonal IgY (Immunoglobulin Yolk) antibodies provides unique and advantageous features that allow highly-specific partitioning of protein mixtures. As a result, previously masked proteins become more accessible for investigation.

The Seppro platform, incorporating SuperMix technology, represents the most complete human protein depletion system available, removing 14 of the most abundant proteins from human serum or plasma, as well as other high and medium abundance proteins. Additional products are available for the depletion of mouse and rat samples, as well as the industry’s only depletion system for the removal of Rubisco.

The ProteoPrep® line of depletion products has been designed for the specific removal of proteins from human serum for further downstream applications. These technologies include material to specifically remove albumin and IgG as well as the ProteoPrep 20 technology for the removal of 20 highly abundant proteins from human plasma/serum (representing 97% of the proteome).

Human Depletion MediaCat. No. Format Size Sample Capacity Proteins DepletedSEP010 Seppro IgY 14 Spin Columns 500 μL resin bed volume 15–20 μL (pooled normal

human plasma or serum)HSA, IgG, Fibrinogen, Transferrin, IgA, IgM, Haptoglobin, α2-Macroglobulin, α1-Acid Glycoprotein, Apo A-I HDL, α1-Antitrypsin, Apo A-II HDL, Complement C3, LDL (Apo B)

SEP020 Seppro IgY14 LC2 6.4 × 63.0 mm (2 mL resin bed volume)

40–50 μL (pooled normal human plasma or serum)

HSA, IgG, Fibrinogen, Transferrin, IgA, IgM, Haptoglobin, α2-Macroglobulin, α1-Acid Glycoprotein, Apo A-I HDL, α1-Antitrypsin, Apo A-II HDL, Complement C3, LDL (Apo B)


100 μL (pooled normal human plasma or serum)



200–250 μL (pooled normal human plasma or serum)


PROTIA ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit

350 μL resin bed volume 25–20 μL (pooled normal human plasma or serum)

Albumin and IgG

PROT20 ProteoPrep 20 Immunodepletion Kit

300 μL resin bed volume 8 μL (pooled normal plasma sample)

Albumin, Transferrin, α1-Acid Glycoprotein, Complement C1q, IgG, Fibrinogen, Ceruloplasmin, Complement C3, IgA, α2-Macroglobulin, Apo A-I HDL, Complement C4, IgM, α1-Antitrypsin, Apo A-II HDL, Plasminogen, IgD, Haptoglobin, Apolipoprotein B, Prealbumin

PROT20LC ProteoPrep 20 Plasma Immunodepletion LC Column

10 mL resin bed volume 100 μL (pooled normal human plasma or serum)

Albumin, Transferrin, α1-Acid Glycoprotein, Complement C1q, IgG, Fibrinogen, Ceruloplasmin, Complement C3, IgA, α2-Macroglobulin, Apo A-I HDL, Complement C4, IgM, α1-Antitrypsin, Apo A-II HDL, Plasminogen, IgD, Haptoglobin, Apolipoprotein B, Prealbumin

http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP010http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP020http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP030http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP040http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/PROTIAhttp://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/PROT20http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/PROT20LC


Seppro DepletionUnmask Low Abundance Proteins

Human Supermix MediaCat. No. Format Size Sample Capacity Proteins DepletedSEP050 Seppro Human Supermix LC2 6.4 × 63 mm (2 mL resin

bed volume)Flow Through volume from IgY14 LC5 Further partitions complex human

plasma/serum samples

SEP060 Seppro Human Supermix LC5 12.7 × 39.5 mm (5 mL resin bed volume)

Flow Through volume from IgY14 LC10 Further partitions complex human plasma/serum samples

Mouse Depletion MediaCat. No. Format Size Sample Capacity Proteins DepletedSEP110 Seppro Mouse Spin Columns 500 μL resin bed volume 15–20 μL (pooled normal mouse plasma

or serum)Mouse serum albumin, IgG, Transferrin, Fibrinogen, IgM, Haptoglobin, α1-Antitrypsin

SEP090 Seppro Mouse LC10 12.7 × 79.0 mm (10 mL resin bed volume)

200–250 μL (pooled normal mouse plasma or serum)

Mouse serum albumin, IgG, Transferrin, Fibrinogen, IgM, Haptoglobin, α1-Antitrypsin

Mouse Supermix MediaCat. No. Format Size Sample Capacity Proteins DepletedSEP100 Seppro Mouse Supermix LC5 12.7 × 39.5 mm (5 mL bed

volume)Flow Through volume from mouse LC10 Further partitions complex mouse

plasma/serum samples

Rat Depletion MediaCat. No. Format Size Sample Capacity Proteins DepletedSEP130 Seppro Rat Spin Columns 500 μL resin bed volume 15–20 μL (pooled normal rat plasma

or serum)Rat serum albumin, IgG, Fibrinogen, Transferrin, IgM, Haptoglobin, α1-Antitrypsin

SEP120 Seppro Rat LC10 12.7 × 79.0 mm (10 mL resin bed volume)

200–250 μL (pooled normal rat plasma or serum)

Rat serum albumin, IgG, Fibrinogen, Transferrin, IgM, Haptoglobin, α1-Antitrypsin

Rubisco Depletion MediaCat. No. Format Size Sample Capacity Proteins DepletedSEP070 Seppro Rubisco Spin Columns 500 μL resin bed volume 15–20 μL (pooled normal plant sample) RuBisCO (Ribulose-1, 5-bisphosphate

carboxylase/oxygenase)

SEP080 Seppro Rubisco LC2 6.4 × 63.0 mm (2 mL resin bed volume)

40–50 μL (pooled normal plant sample) RuBisCO (Ribulose-1, 5-bisphosphate carboxylase/oxygenase)

http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP050http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP060 http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP110http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP090http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP100http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP130http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP120 http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP070 http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/SEP080

biomapping

Seppro DepletionIncorporating SuperMix Technology

The Seppro platform, incorporating SuperMix technology, represents the most complete human depletion system available, removing 14 of the most abundant proteins from human serum or plasma, as well as other high (HAP) and medium abundance proteins (MAP). This system consists of the use of the IgY14 depletion column (LC5 or LC10) in conjunction with the SuperMix column (LC2 or LC5). The IgY SuperMix is generated by immunizing chickens with human plasma proteins from which 12 HAP have been removed. The majority of resultant antibodies incorporated into the depletion resin are against MAP and/or immunoreactive proteins.

Dynamic Range of Human Plasma

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Interleukins, etc

0 1 2 3 4 5 6 7 8 9 10 11 12

Classical Plasm

a Protein

s

Normal Range Abundance Log10

Concentration in pg/ml

Protein Mass (Percentage)

HAP

MAP

Sample(250 ml)

IgY14

SuperMix

MAP Medium-AbundantProteins

Eluate (E1)HAP 14 Highly- Abundant

Proteins

Eluate (E2)

Flow-Through (F1)

HemoglobinAlbuminIgG Total

TransferrinFibrinogen

IgA TotalAlpha-2-Macroglobulin

IgM TotalAlpha-1-Antitrypsin

C3 ComplementHaptoglobin

Apolipoprotein A-1Apolipoprotein B

Alpha-1-acid GlycoproteinLipoprotein(a)

Factor HCeruloplasmin

C4 ComplementComplement Factor B

PrealbuminC9 Complement

C1q ComplementC8 Complement

C5 Complement Plasminogen

IgDC1 inhibitor

C6 ComplementC7 Complement

Complement Factor IRetinol Binding Protein

iC3bThyroxin Binding Globulin

C2 Complement ProteinThrombus Precursor Protein

C-reactive ProteinBb Fragment

C3a Complement ProteinFerritinRantes

SC5b-9 ComplexMyoglobin

ThyroglobulinTPA

C5a ComplementNeuron-Speci�c Enolase

C-PeptideAlpha-fetoprotein

TNF-Binding ProteinsProstate-Speci�c Antigen

Prostatic Acid PhosphataseCEA

Myelin Basic ProteinTroponin I

Interleukin-1raMIP-1 betaTroponin T

Interleukin-8MIP-1 alpha

Tissue FactorGCSF

Interferon AlphaInterleukin-2Interleukin-4

TNF-AlphaInterferon GammaInterleukin-1 Beta

Interleukin-12Interleukin-10

Interleukin-5Interleukin-6

LAP

LAP

Results of SuperMix ColumnsM P F1 E2 F2

MW (kDa)

250150100

75

50

37

252015

M: MW Marker

P: Plasma

F1: Flow-Through of IgY14

E2: Bound/Eluted Fraction from SuperMix

F2: Flow-Through of IgY14 + IgY-SuperMix

4-20% SDS-PAGE under reducing conditions

4 mg Protein loaded per lane

Flow-Through (F2)

Tissue Leakag

e

Seppro Human IgY14/SuperMixAdd Focus to Your Protein Biomarker Research

sigma.com/seppro

http://sigma.com/seppro


Custom AQUA PeptidesAccurate Mass Spectrometric Quantitation

AQUA Peptides™Focus Your Protein Biomarker Research

The AQUA TechnologyDeveloped in collaboration with the Protein Quantitation Consortium, Sigma’s AQUA Peptides™ enable accurate, efficient mass spectrometric quantitation of protein biomarkers.

• Focus your analysis on significant protein biomarkers• Accurately quantitate low abundance proteins• Eliminate costly, time consuming stable isotope labeling steps• Measure site-specific phosphorylation states• Validate gene silencing at the protein level

Custom AQUA PeptidesTo meet the specific demands of AQUA experimentation, Sigma has developed a specialized custom peptide offering. Custom AQUA Peptides are synthesized using fully labeled 98 atom % 13C and

98 atom % 15N enriched amino acids (one labeled amino acid per peptide) and are stringently tested to ensure high purity (HPLC), accurate molecular mass (ESI-MS), and specific peptide content.

• >95% pure by HPLC• Quantitated by AAA • Confirmed by ESI-MS• 5 × 1 nmole packages

Ordering InformationProduct Name Package SizeCustom AQUA Peptide 5 × 1 nmole

To order Custom AQUA Peptides, call 1-800-325-3010 or visit, sigma.com/aquaorder

PEPscreen®An Enabling Technology for Rapid Characterization and Selection of MS Compatible Peptides

To order, call 800-234-5362 or visit, sigma.com/pepscreen

Sigma’s proprietary peptide synthesis platform enables fast and efficient high-throughput parallel synthesis of sub-milligram quantities of peptides.

A Perfect Solution for:• Qualifying peptides for mass spectrometry suitability regarding

chromatographic, ion suppression, and transition states

• Screening multiple proteotypic peptides quickly and economically• Identifying the best (isotopically labeled) AQUA Peptide™ candidate

for optimal protein quantitation performance using your analytical techniques

PEPscreen Product Specifications• Quantity: 0.5–2 mg – Gross weight provided for all peptides• Length: 6–20 mers• Modifications available • Format: dried in 96-tube rack format• QC: MALDI-ToF MS performed on all peptides — Electronic PDF files

of QC data and peptide directory supplied on CD

• C-terminal acid or amide• Minimum order size is 24 peptides

http://sigma.com/aquaorderhttp://sigma.com/pepscreen

biomapping

StandardizeWith UPS from Sigma

Sigma offers both the Universal Proteomics Standard and the Proteomics Dynamic Range Standard as complex, well-defined, well-characterized reference standards for mass spectrometry. Both standards contain the same 48 human proteins ranging in molecular mass from 6,000 to 83,000 Daltons. Each constituent protein has been HPLC purified and AAA quantitated prior to formulation.

• Troubleshoot and optimize your analytical protocol• Confirm system suitability before analyzing critical samples• Normalize analytical results day-to-day or lab-to-lab• Determine your limit of detection

Universal Proteomics StandardUPS1

Developed in collaboration with the Association of Biomolecular Resource Facilities (ABRF) Proteomics Standards Research Group (sPRG), the Universal Proteomics Standard contains 48 human proteins (5 pmoles of each) ranging in molecular mass from 6,000 to 83,000 Daltons.

Proteomics Dynamic Range StandardUPS2

This standard is an enhancement of Sigma’s original Universal Proteomics Standard (UPS1). The same complex mixture of 48 human proteins has been formulated into a dynamic range of concentration levels, ranging from 50 pmoles to 0.5 fmoles.

Ordering Information

Universal Proteomics Standard (UPS) Standardize Your Proteomics Research

To learn more, visit sigma.com/ups

Cat. No. Product NameUPS1 Universal Proteomics Standard SetUPS2 Proteomics Dynamic Range Standard Set

Criteria Verified ByProteins More than 175 proteins were screened by SDS-PAGE

and LC/MS to be considered suitable for use in the standard

Minimal PTMs LC/MS of the intact proteins to verify homogeneity of the protein population

Diversity Proteins are of wide-ranging molecular masses, hydrophobicities, isoelectric points, etc.

High Purity Proteins are of wide-ranging molecular masses, hydrophobicities, isoelectric points, etc.

Accurate Quantitation Amino Acid Analysis in triplicate

Table 1: Criteria used to develop and produce the Universal Proteomics Standards line of products.

http://sigma.com/upshttp://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/UPS1http://www.sigma-aldrich.com/catalog/search/ProductDetail/SIGMA/UPS2


The Universal Proteomics Standard (UPS1) and the Proteomics Dynamic Range Standard (UPS2) contain the following proteins:UniProt Accession Number3

UPS1 Amount

(fmol)

UPS2 Amount

(fmol) UniProt Protein Name (Synonym)MW (Da)

CalculatedSource or

recombinant Host Tag PTMs Potential

P00915 5,000 50,000 Carbonic anhydrase 1 28,738 Erythrocytes — — Acetylation

P00918 5,000 50,000 Carbonic anhydrase 2 29,095 Erythrocytes — — Acetylation

P01031 5,000 50,000 Complement C5 [Complement C5a] 8,266 Recombinant E. coli — —

P69905 5,000 50,000 Hemoglobin alpha chain 15,127 Erythrocytes — — —

P68871 5,000 50,000 Hemoglobin beta chain 15,867 Erythrocytes — — Acetylation, Nitrosylation, Glycosylation

P41159 5,000 50,000 Leptin 16,024 Recombinant E. coli — —

P02768 5,000 50,000 Serum albumin 66,393 Recombinant Pichia pastoris — —

P62988 5,000 50,000 Ubiquitin 9,387 Recombinant E. coli 6-His —

P04040 5,000 5,000 Catalase 59,583 Erythrocytes — — —

P00167 5,000 5,000 Cytochrome b5 16,021 Recombinant E. coli 6-His —

P01133 5,000 5,000 Epidermal growth factor 6,211 Recombinant E. coli — —

P02144 5,000 5,000 Myoglobin 17,051 Heart — — —

P15559 5,000 5,000 NAD(P)H dehydrogenase [quinone] 1 [DT Diaphorase] 30,984 Recombinant E. coli — —

P62937 5,000 5,000 Peptidyl-prolyl cis-trans isomerase A [Cyclophilin A] 17,947 Recombinant E. coli — —

Q06830 5,000 5,000 Peroxiredoxin 1 22,106 Recombinant E. coli — —

P63165 5,000 5,000 Small ubiquitin-related modifier 1 [SUMO-1] 37,420 Recombinant E. coli GST —

P00709 5,000 500 Alpha-lactalbumin 14,070 Milk — — Glycosylation

P06732 5,000 500 Creatine kinase M-type [CK-MM] 43,070 Heart — — —

P12081 5,000 500 Histidyl-tRNA synthetase [Jo-1] 58,223 Recombinant E. coli — —

P61626 5,000 500 Lysozyme C 14,692 Milk — — —

Q15843 5,000 500 Neddylin [Nedd8] 9,071 Recombinant E. coli — —

P02753 5,000 500 Retinol-binding protein 21,065 Urine — — —

P16083 5,000 500 Ribosyldihydronicotinamide dehydrogenase (quinone) [Quinone oxidoreductase 2 or NQO2]

25,817 Recombinant E. coli — —

P63279 5,000 500 Ubiquitin-conjugating enzyme E2 I [UbcH9] 17,995 Recombinant E. coli — —

P01008 5,000 50 Antithrombin-III 49,033 Plasma — — Glycosylation

P61769 5,000 50 Beta-2-microglobulin 11,729 Urine — — —

P55957 5,000 50 BH3 interacting domain death agonist [BID] 21,978 Recombinant E. coli — —

O76070 5,000 50 Gamma-synuclein 15,363 Recombinant E. coli 6-His —

P08263 5,000 50 Glutathione S-transferase A1 [GST A1-1] 25,482 Recombinant E. coli — —

P01344 5,000 50 Insulin-like growth factor II 7,464 Recombinant E. coli — —

P01127 5,000 50 Platelet-derived growth factor B chain 12,286 Recombinant E. coli — —

P10599 5,000 50 Thioredoxin 12,424 Recombinant E. coli 6-His —

P01579 5,000 5 Interferon gamma (IFN-gamma) 16,879 Recombinant E. coli — —

P99999 5,000 5 Cytochrome c [Apocytochrome c] 11,608 Recombinant E. coli — —

P06396 5,000 5 Gelsolin 8 2,954 Plasma — — Phosphorylation

P09211 5,000 5 Glutathione S-transferase P [GST] 23,220 Placenta — — —

P01112 5,000 5 GTPase HRas [Ras protein] 21,292 Recombinant E. coli — —

P02787 5,000 5 Serotransferrin [Apotransferrin] 75,143 Plasma — — Glycosylation

O00762 5,000 5 Ubiquitin-conjugating enzyme E2 C [UbcH10] 20,473 Recombinant E. coli 6-His —

P51965 5,000 5 Ubiquitin-conjugating enzyme E2 E1 [UbcH6] 22,222 Recombinant E. coli 6-His —

P08758 5,000 0.5 Annexin A5 35,782 Placenta — — Acetylation

P02741 5,000 0.5 C-reactive protein 23,030 Plasma — — —

P05413 5,000 0.5 Fatty acid-binding protein 14,716 Plasma — — Acetylation, Phosphorylation

P10145 5,000 0.5 Interleukin-8 8,381 Recombinant — E. coli —

P02788 5,000 0.5 Lactotransferrin 78,289 Milk — — Glycosylation

P10636 5,000 0.5 Microtubule-associated protein tau [Tau protein] 46,810 Recombinant E. coli 6-His —

P00441 5,000 0.5 Superoxide dismutase [Cu-Zn] 15,800 Erythrocytes — — Acetylation

P01375 5,000 0.5 Tumor necrosis factor [TNF-alpha] 17,350 Recombinant E. coli — —

biomapping

Panorama Antibody Array TechnologyScreen Expression Changes in Proteins Faster

The Panorama XPRESS Profiler725 contains 725 highly specific antibodies, arrayed in duplicate on nitrocellulose slides compatible with the majority of DNA array scanners available today.

The array antibodies detect human, mouse, and rat proteins associated with a variety of key cell signaling and gene regulation pathways. Targeted-content subsets of antibodies to Mitogen-Activated Protein Kinase (MAPK), Protein Kinase C (PKC), and p53 protein families are also represented on the array.

Research of biological systems from a genomic perspective focuses on mRNA expression levels and high-throughput data generation was accelerated by the advent of DNA arrays. However, mRNA undergoes a number of alternative processing steps prior to and following translation.

There is often poor correlation between mRNA and protein expression, and this has driven development of antibody-based technologies that enable measurement of biologically-relevant changes at the proteome level.

With the sensitivity of antibody detection, Panorama Antibody Array technology enables researchers to screen expression changes in large numbers of proteins in significantly less time than it would take using conventional protein detection methodology.

The Panorama Antibody Array XPRESS Profiler725 is a high-throughput tool for multiplex protein expression profiling of serum, cell or tissue extracts.

Protein Expression Profiling... broad and fast• Use serum, cell, or tissue samples• Rapid, multiplex analysis of protein expression• Profile 725 proteins in a single assay• Antibodies react with human, mouse, and rat proteins• Compatible with Cy™3/Cy5 detection

Panorama® Antibody ArraysYour Key to Fast Protein Expression Profiling

Step 2:Label samples with Cy3/Cy5

and mix

Step 3:Incubate on the array

Step 4:Scan the array

Sample BSample A

Step1:Extract proteins

How it Works

Applications• Biomarker discovery and drug development• Identify gene silencing (RNAi) off-target effects• Cell response to external stimuli (e.g., starvation, growth factors,

serum, drugs)

• Differential protein expression in tissue samples (e.g, heart, brain, lung etc.)


The Panorama Antibody Array XPRESS Profiler725Delivers Multiplex Protein Expression Profiling of Serum, Cell or Tissue Extracts

Kit Contents• Panorama Antibody Slides (2 Each)• quadriPERM® Cell Culture Vessels (2 Each)• Extraction/Labeling Buffer• Protease Inhibitor Cocktail• Phosphatase Inhibitor Cocktail II• Benzonase®, Ultrapure• Array Incubation Buffer• Phosphate Buffered Saline, pH 7.4, with TWEEN® 20 (Washing Buffer)• SigmaSpin™ Post-Reaction Clean-Up Columns (8 each)• Collection Tubes, Polypropylene, 2 mL (16 each)• Panorama Antibody List• Slide Analysis Software Diskette

Differential Expression Between Two Cell StatesExpression of proteins in F9 mouse stem cells induced to differentiate by retinoic acid. The total cell extract of F9 cells non-treated or treated for 4 days with retinoic acid were prepared and labeled with Cy3 and Cy5, respectively. Equal amounts of Cy3 or Cy5 labeled protein were incubated on the slide. The experiment was repeated at least three times.

Panorama Antibody Array technology enables researchers to screen expression changes in large numbers of proteins, in significantly less time than it would take using conventional protein detection methods.

PolyglutamylatedTubulin

ß-Tubulin I

Unchanged

ß-Tubulin IV

Connexin 43

Up-Regulated Down-Regulated

PAR4

Grb2

Non-Treated (Cy3) Treated (Cy5)

– +

– + – +

– +

– +

– +

MPJ838351075

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New Levels of Accuracy for All Your Proteomics Research · 2016. 2. 28. · SEP010 Seppro IgY 14 Spin Columns 500 μL resin bed volume 15–20 μL (pooled normal human plasma or serum)