Product Pathways - Apoptosis / Autophagy
Pro-Survival Bcl-2 Family Antibody Sampler Kit (Human Specific) #9941
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|---|---|
| Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb # 2827 | 40 microliters | W IF-IC F | H | 28 | Rabbit |
| Phospho-Bcl-2 (Thr56) Antibody (Human Specific) # 2875 | 40 microliters | W | H | 28 | Rabbit |
| Bcl-2 (50E3) Rabbit mAb # 2870 | 40 microliters | W IP | H M R Mk (C) (B) (Dg) | 26 | Rabbit |
| Bcl-xL (54H6) Rabbit mAb # 2764 | 40 microliters | W IP IHC-P IHC-F F | H M R Mk | 30 | Rabbit |
| Mcl-1 Antibody # 4572 | 40 microliters | W | H | 40 | Rabbit |
| Anti-rabbit IgG, HRP-linked Antibody # 7074 | 100 microliters | Goat |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IHC-F=Immunohistochemistry (Frozen)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
C=Chicken
B=Bovine
Dg=Dog
Specificity / Sensitivity
Each antibody in the Pro-Survival Bcl-2 Family Antibody Sampler Kit (Human Specific) recognizes only its specific target. The antibodies do not cross-react with other Bcl-2 family members.
Western Blotting
Western blot analysis of extracts from Jurkat and HeLa (human), COS (monkey), NIH/3T3 and L929 (mouse), and PC12 and C6 (rat) cells, using Bcl-xL (54H6) Rabbit mAb #2764.
Western Blotting
Western blot analysis of extracts from THP-1 cells, untreated or nocadazole-treated (1.0 µg/mL) overnight, using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb #2827 (upper), Phospho-Bcl-2 (Thr56) Antibody (Human Specific) #2875 (middle) and Bcl-2 (50E3) Rabbit mAb #2870 (lower).
Western Blotting
Western blot analysis of extracts from various cell types using Bcl-2 (50E3) Rabbit mAb #2870.
Western Blotting
Western blot analysis of extracts from Raji, Ramos, and BL41-3 (a subline of BL41 Burkitt lymphoma cells found to have amplified expression of Mcl-1) cells, using Mcl-1 Antibody #4572.
Flow Cytometry
Flow cytometric analysis of Jurkat cells, using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb #2827 versus propidium iodide (DNA content).
IF-IC
Immunofluorescent staining of MCF-7 cells with or without treatment of paclitaxel (1 μM, overnight), using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb #2827.
Source / Purification
Monoclonal antibodies were produced from rabbits immunized with synthetic peptides (KLH-coupled) corresponding to residues surrounding phospho-Ser70 of human Bcl-2, Asp61 of human Bcl-xL, and residues at the carboxy terminus of Bcl-2. Polyclonal antibodies are produced by immunizing rabbits with synthetic peptides (KLH-coupled) corresponding to residues surrounding phospho-Thr56 of human Bcl-2 and Ser121 of human Mcl-1. Antibodies are purified by protein A and peptide affinity chromatography.
Background
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.
Several phosphorylation sites have been identified within Bcl-2 including Thr56, Ser70, Thr74 and Ser87 (6). These phosphorylation sites may be targets of the ASK1/MKK7/JNK1 pathway, and phosphorylation of Bcl-2 may be a marker for mitotic events (7,8). Mutation of Bcl-2 at Thr56 or Ser87 inhibits its anti-apoptotic activity during glucocorticoid-induced apoptosis of T lymphocytes (9). Interleukin 3 and JNK-induced Bcl-2 phosphorylation at Ser70 may be required for its enhanced antiapoptotic functions (10).
- Cory, S. et al. (2003) Oncogene 22, 8590-8607.
- Antonsson, B. and Martinou, J. (2000) Exp. Cell Res. 256, 50-57.
- Sharpe, J.C. et al. (2004) Biochim. Biophys. Acta. 1644, 107-113.
- Korsmeyer, S.J. et al. (1993) Semin. Cancer Biol. 4, 327-337.
- Bouillet, P. and Strasser, A. (2002) J. Cell Sci. 115, 1567-1574.
- Maundrell, K. et al. (1997) J. Biol. Chem. 272, 25238-25242.
- Yamamoto, K. et al. (1999) Mol. Cell Biol. 19, 8469-8478.
- Ling, Y. H. et al. (1998) J. Biol. Chem. 273, 18984-18991.
- Huang, S.J. and Cidlowski, J.A. (2002) FASEB 16, 825-832.
- Deng, X. et al. (2001) J. Biol. Chem. 276, 23681-23688.
Application References
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