The Bcl-2 family consists of a number of evolutionarily conserved proteins containing Bcl-2 homology domains (BH) that regulate apoptosis through control of mitochondrial membrane permeability and release of cytochrome c (1-3). Four BH domains have been identified (BH1-4), which mediate protein interactions. The family can be separated into three groups based upon function and sequence homology: pro-surivival members including Bcl-2, Bcl-xL, Mcl-1, A1 and Bcl-w; pro-apoptotic proteins including Bax, Bak and Bok, and "BH3 only" proteins Bad, Bik, Bid, Puma, Bim, Bmf, Noxa and Hrk. Interactions between death-promoting and death-suppressing Bcl-2 family members has led to a rheostat model in which the ratio of pro-apoptotic and anti-apoptotic proteins controls cell fate (4). Thus, pro-survival members exert their behavior by binding to and antagonizing death-promoting members. In general, the "BH3-only members" can bind to and antagonize the pro-survival proteins leading to increased apoptosis (5). While some redundancy of this system likely exists, tissue specificity, transcriptional and post-translational regulation of many of these family members can account for distinct physiological roles.
Bad is a pro-apoptotic member of the Bcl-2 family that can displace Bax from binding to Bcl-2 and Bcl-xL, resulting in cell death (6,7). 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 (7). Phosphorylation at these sites results in the binding of Bad to 14-3-3 proteins and the inhibition of Bad binding to Bcl-2 and Bcl-xL (7). Akt has been shown to promote cell survival via its ability to phosphorylate Bad at Ser136 (8,9). Ser112 has been shown to be the substrate in vivo and in vitro of p90RSK (10,11) and mitochondria-anchored PKA (12).
This kit contains enough primary and secondary antibodies to perform four Western mini-blot experiments.
B0807-02C1: Bad phospho Mab (Ser112) Mo x Mo 1x40ul
B0807-02C2: Bad Pab Antibody Rabbit x Mo 1x40ul
B0807-02C3: Bax Pab Antibody Rabbit x Mo 1x40ul
B0807-02C4: Bik Pab Antibody Rabbit x Mo 1x40ul
B0807-02C5: Bim Pab Antibody Rabbit x Mo 1x40ul
B0807-02C6: Bmf Pab Antibody Rabbit x Mo 1x40ul
B0807-02C7: Bok Pab Antibody Rabbit x Mo 1x40ul
B0807-02C8: Puma Pab Antibody Rabbit Mo 1x40ul
B0807-02C9: IgG Pab (HRP) Goat x Mo 1x100ul
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.