Bad is a proapoptotic member of the Bcl-2 family that can displace Bax from binding to Bcl-2 and Bcl-XL, resulting in cell death (1,2). Survival factors such as IL-3 can inhibit the apoptotic activity of Bad by activating intracellular signaling pathways that result in the phosphorylation of Bad at Ser112 and Ser136 (2). Phosphorylation at these sites results in the binding of Bad to 14-3-3 proteins and the inhibition of Bad binding to Bcl-XL and Bcl-2 (2). Akt has been shown to promote cell survival via its ability to phosphorylate Bad at Ser136 (3,5). Ser112 has been shown to be the substrate in vivo and in vitro of p90RSK (6,7) and mitochondria-anchored PKA (8).
Nonphosphorylated Bad Control Proteins: Nonphosphorylated Bad peptide fusion serves as a negative control. Supplied in SDS Sample Buffer.
Phosphorylated Bad Control Proteins: Phosphorylated Bad peptide fusion serves as a positive control.
Directions for Use:
As controls, we recommend using 10ul of phosphorylated and nonphosphorylated Bad control proteins. Boil for 2 minutes prior to use. 100ul wil be sufficient for 10 blots.