HOW DID OUR ANTIBODY PERFORM?

ANTIBODY CUSTOMER REVIEW: Phospho-PP1α (T320) Polyclonal Antibody (STJ91134)

What was used?Primary antibody: STJ91134 Phospho-PP1α (T320)

Provider: St John's Laboratory

Dilution ratio: 1:500

Application: Western Blot

Materials for validation: HEK cell line

Click for product data sheet PDF

What Was The Protocol?Treatment of Materials Cells were cultured to full confluence plating density. Lysis occurred with protease inhibitors

(Roche), 1% triton, 50 mM Tris-HCL, 150 mM NaCl and a phosphatase inhibitors cocktail (Sigma). 5X of sample buffer was then added.

Gel Electrophoresis 15% polyacrylamide gel, constant voltage 160 V, for 60 minutes.

Transfer 0.45 µm PVDF membrane. 1 piece of gel (semi-dry transfer), constant current 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, 4˚C and left overnight.

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

Secondary Antibody Probing Secondary antibody was added to the solution of 1X TBST with 5% BSA with a 1:2000 dilution ratio, for 1.5 hours.

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

Visualization Visualization: ECL visualization for 3 min (the visualization time was determined by actual result. It was ensured that the visualization time was identical for the same antigen).

Gel electrophoresis information: 15% polyacrylamide gel, constant voltage 160 V, 60 min.

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

No. Antigen Loading amount Primary antibody

Primary antibody

dilution ratio

Secondaryantibody dilution

ratioTarget band KD Visualization time

1 Hek lysate 40ug STJ91134 phospho PP1 1:500 1:2000 ~37KD 60S

What were the results?

What did the customer think?

Antibody Specificity:

Antibody Rating:

Testing results were provided independently by S. Hilfiker at Instituto de Parasitología y Biomedicina "López - Neyra".

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