Products:Cell Biology >> Apoptosis >> Intracellular >> Associated Proteins
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Predicted band size : 17 kDa
Lysates prepared from Hek293T cells transfected with rat WT Bim alone (1, 4), cotransfected with rat WT Bim and activated MEKK1 (2, 5) or with
mutant S65A Bim and MEKK1 (3, 6), were resolved by SDS-PAGE on a 14% polyacrylamide gel and transferred to PVDF. Membranes were blocked with 3% Milk-TBST buffer for one hour at room temperature, and incubated with Bim Pan antibody (1, 2, 3) or ab17935 (4, 5, 6) for two hours at room temperature in 3% Milk-TBST buffer. After washing, membranes were incubated with a goat F(ab')2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal™ method.
The data show that the signal is detected only in lysates co-expressing rat WT Bim and active MEKK1. This signal is abolished in cells expressing mutant rat Bim S65A and MEKK1, verifying that the signal is site-phosphospecific.
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