MMP2 (matrix metalloproteinase 2) is a collagenase that is involved in a vast number of processes including angiogenesis, tumour invasion and inflammation. It is encoded by the MMP2 gene in humans. The protein is known to have associations with diseases such as Torg-Winchester syndrome and arthritis.

1. HOW DID OUR
ANTIBODY PERFORM?
ANTIBODY CUSTOMER REVIEW: MMP2 Polyclonal Antibody (STJ28003)

2. 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

3. 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.

4. 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
Secondary
antibody
dilution ratio
Target band KD
Visualization
time
1 hCSF lysate 50µg
STJ96177.
RARA antibody 1:1000 1:5000 ~68KD N/A

5. What were the results?

6. 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.”

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