TNFa was immunoprecipitated using:
Lane A:0.5 mg Hela Whole Cell Lysate
Lane B:0.5 mg HepG2 Whole Cell Lysate
Lane C:0.5 mg SW480 Whole Cell Lysate2 µL anti-TNFa rabbit polyclonal antibody and 15 μl of 50 % Protein G agarose.Primary antibody:
Anti-TNFa rabbit polyclonal antibody,at 1:200 dilutionSecondary antibody:
Dylight 800-labeled antibody to rabbit IgG (H+L), at 1:5000 dilutionDeveloped using the odssey technique.
Performed under reducing conditions.Predicted band size: 23 kDa
Observed band size: 33 kDa
Anti-TNF-alpha rabbit polyclonal antibody at 1:500 dilution
Lane A: HepG2 Whole Cell LysateLysates/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:25 kDa
Observed band size:27 kDa