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HOW DID OUR ANTIBODY PERFORM?
ANTIBODY CUSTOMER REVIEW: MMP2 Polyclonal Antibody (STJ28003)
What was used?Primary antibody: STJ28003 MMP2 Antibody
Provider: St John's Laboratory
Dilution ratio: 1:1000
Application: Western Blot
Materials for validation: Human primary corneal stromal fibroblasts (hCSF)
Click for product data sheet PDF
What Was The Protocol?Treatment of Materials When cells were cultured to 70%-80% plating density, lysis buffer with protease inhibitors
(Roche) and equal volume of 2X loading buffer were added for 5 minutes at 100˚C.
Gel Electrophoresis 10% Polyacrylamide gel with a constant voltage of 180V for 60 minutes.
Transfer 0.2 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100V for 60 minutes.
Blocking 1X TBST with 5% BSA for 1 hour.
Primary Antibody Probing Primary antibody was added to the solution of 1X TBST with 5% BSA, RT for 4 hours.
Membrane Wash Membrane was washed with 1X TBST 4 times, for 5 minutes each .
Secondary Antibody Probing Secondary antibody was added to the solution of 1X TBST with 5% BSA with a 1:5000 dilution ratio, for 2 hours.
Membrane Wash Membrane was washed with 1X TBST 4 times, for 5 minutes each .
Visualization Li-Cor Odyssey IF scanner at maximum resolution.
What was the protocol?Gel electrophoresis information: 10% polyacrylamide gel, constant voltage of 180V for 60 minutes.
Transfer information: 0.2 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100V for 60 minutes.
No. Antigen Loading amount
Primary antibody
Primary antibody
dilution ratio
Secondaryantibody
dilution ratioTarget band KD Visualization
time
1 hCSF lysate 50µgSTJ96177.
RARA antibody 1:1000 1:5000 ~68KD N/A
What were the results?
What did the customer think?
Antibody Specificity:
Antibody RPPA Rating:
Testing results were provided independently by R. Gouveia at Newcastle University. They stated “The antibody reveals good specificity with a clear background.”
JOIN THE ANTIBODY VALIDATION PROJECT
1. Order any STJ9 catalogued product using this online form.
2. Test it in your laboratory using your preferred method.
3. Return a fully completed review form to receive rewards.
…in 3 simple steps…
@StJohnsLabs
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