HOW DID OUR ANTIBODY PERFORM?

ANTIBODY CUSTOMER REVIEW: RARA Polyclonal Antibody (STJ26160 )

What was used?Primary antibody: STJ26160 RARA 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 U87 cell lysate 50µgSTJ96177.

RARA antibody 1:1000 1:5000 ~48KD 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 reasonable specificity with good a background. Additional bands might constitute conjugated products not dissociated by PAGE conditions.”

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