AKR1B1 was immunoprecipitated using:
Lane A:0.5 mg Jurkat Whole Cell Lysate
Lane B:0.5 mg Hela Whole Cell Lysate
Lane C:0.5 mg A431 Whole Cell Lysate2 µL anti-AKR1B1 rabbit polyclonal antibody and 15 μl of 50 % Protein G agarose.Primary antibody:
Anti-AKR1B1 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: 35 kDa
Observed band size: 35 kDa
Anti-AKR1B1 rabbit polyclonal antibody at 1:500 dilution
Lane A: Hela Whole Cell Lysate
Lane B: MCF-7 Whole Cell Lysate
Lane C: Jurkat Whole Cell Lysate
Lane D: 293T 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:36 kDa
Observed band size:34 kDa
(We are unsure as to the identity of these extra bands.)