P4HB was immunoprecipitated using:
Lane A:0.5 mg MCF-7 Whole Cell Lysate2 µL anti-P4HB rabbit monoclonal antibody and 15 μl of 50 % Protein G agarose.Primary antibody:
Anti-P4HB rabbit monoclonal antibody,at 1:200 dilutionSecondary antibody:
Clean-Blotô IP Detection Reagent (HRP) at 1:1000 dilutionDeveloped using the DAB staining technique.
Performed under reducing conditions.Predicted band size: 57 kDa
Observed band size: 57 kDa
Anti-P4HB rabbit monoclonal antibody at 1:100 dilution
Lane A: MCF7 Whole Cell Lysate
Lane B: HepG2 Whole Cell Lysate
Lysates/proteins at 30 μg per lane.
Goat Anti-Rabbit IgG H&L (Dylight800) at 1/10000 dilution.
Developed using the Odyssey technique.
Performed under reducing conditions.
Predicted band size:57 kDa
Observed band size:57 kDa
(We are unsure as to the identity of these extra bands.)