HOW DID OUR ANTIBODY PERFORM?

ANTIBODY CUSTOMER REVIEW: β-actin Monoclonal Antibody (STJ96941)

What was used?Primary antibody: STJ96941 β-actin Antibody

Provider: St John's Laboratory

Dilution ratio: 1:5000

Application: Western Blot

Materials for validation: HEK293 cell lysates

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 and

equal volume of 2X loading buffer were added for 10 minutes in a water bath at 100˚C.

Gel Electrophoresis 12% Polyacrylamide gel with a constant voltage of 185V for 60 minutes.

Transfer 0.45 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 minutes.

Blocking 1X TBST with 5% skimmed milk powder was used and left overnight at 4°C.

Primary Antibody Probing Primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder and left at room temperature for 1 hour.

Membrane Wash Membrane was washed 3 times with 1X TBST.

Secondary Antibody Probing Secondary antibody was added to the solution of 1X TBST with 5% skimmed milk powder with a 1:5000 dilution ratio and left at room temperature for 1 hour.

Membrane Wash Membrane was washed 3 times with 1X TBST.

Visualization ECL visualization for 5 minutes (the visualization time was determined by actual result. It was ensured that the visualization time was identical for the same antigen).

What was the protocol?Gel electrophoresis information: 12% Polyacrylamide gel with a constant voltage of 185V for 60 minutes.

Transfer information: 0.45 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 minutes.

No. Antigen Loading amount

Primary antibody

Primary antibody

dilution ratio

Secondaryantibody

dilution ratioTarget band KD Visualization

time

1 HEK293 lysate 10ul STJ96941 β-actin antibody 1:5000 1:5000 ~43KD N/A

What were the results?

β-actin Antibody

Predicted Band: 43 kDa Actual Band: 43 kDa

What did the customer think?

Antibody Specificity:

Antibody Rating:

Testing results were provided independently by Viva Biotech Ltd. They stated “Satisfied! The antibody works in accordance with expectation.”

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