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9941
Pro-Survival Bcl-2 Family Antibody Sampler Kit

Pro-Survival Bcl-2 Family Antibody Sampler Kit #9941

Western Blotting Image 1

Western blot analysis of extracts from Jurkat cells, untreated or treated with paclitaxel (1 μM, overnight) and with or without λ phosphatase, using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb (upper) or Bcl-2 #2876 (lower).

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Western Blotting Image 2

Western blot analysis of Jurkat cells, untreated or treated with paclitaxel for the indicated times, using Phospho-Bcl-2 (Thr56) Antibody (Human Specific) (top) or Bcl-2 Antibody (Human Specific) #2872 (bottom).

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Western Blotting Image 3

Western blot analysis of extracts from various cell lines using Bcl-2 (D55G8) Rabbit mAb (Human Specific).

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Western Blotting Image 4

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.

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Western Blotting Image 5

Western blot analysis of extracts from various cell lines using Mcl-1 (D35A5) Rabbit mAb.

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Western Blotting Image 6

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Flow Cytometry Image 7

Flow cytometric analysis of Jurkat cells, using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb versus propidium iodide (DNA content).

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Western Blotting Image 8

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® BcL-xL siRNA I (+) or SignalSilence® Bcl-xL siRNA II #6363 (+), using Bcl-xL (54H6) Rabbit mAb #2764 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The Bcl-xL (54H6) Rabbit mAb confirms silencing of Bcl-xL expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

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Western Blotting Image 9

Western blot analysis of extracts from 293T cells, mock transfected or transfected with human or mouse Mcl-1 constructs, using Mcl-1 (D35A5) Rabbit mAb.

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IF-IC Image 10

Confocal immunofluorescent analysis of SH-SY5Y cells using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb (red) and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IP Image 11

Immunoprecipitation of Bcl-xL from Jurkat cell extracts, using Bcl-xL (54H6) Rabbit mAb. Lane 1 is the lysate control, lane 2 is antibody alone and lane 3 is antibody plus lysate.

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IHC-P (paraffin) Image 12

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Bcl-xL (54H6) Rabbit mAb.

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IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded 4TI syngeneic mouse tumor, using Bcl-xL (54H6) Rabbit mAb # 2764.

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IHC-P (paraffin) Image 14

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Bcl-xL (54H6) Rabbit mAb in the presence of control peptide (left) or Bcl-xL Blocking Peptide #1225 (right).

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IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, showing cytoplasmic localization, using Bcl-xL (54H6) Rabbit mAb.

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IHC-F (frozen) Image 16

Immunohistochemical analysis of frozen H1650 xenograft, showing cytoplasmic localization using Bcl-xL (54H6) Rabbit mAb.

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Flow Cytometry Image 17

Flow cytometric analysis of untreated Jurkat cells, using Bcl-xL (54H6) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IF-IC Image 18

Confocal immunofluorescent analysis of HeLa cells using Bcl-xL (54H6) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb 2827 20 µl
  • WB
  • IF
  • F
H 28 Rabbit IgG
Phospho-Bcl-2 (Thr56) Antibody (Human Specific) 2875 20 µl
  • WB
H 28 Rabbit 
Bcl-2 (D55G8) Rabbit mAb (Human Specific) 4223 20 µl
  • WB
  • IP
H 26 Rabbit IgG
Bcl-xL (54H6) Rabbit mAb 2764 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Mk 30 Rabbit IgG
Mcl-1 (D35A5) Rabbit mAb 5453 20 µl
  • WB
H M Mk 40 (human), 35 (mouse) Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Pro-Survival Bcl-2 Family Antibody Sampler Kit provides an economical means to examine several members of the Bcl-2 family. The kit contains enough primary and secondary antibodies to perform two western blot experiments.

Each antibody in the Pro-Survival Bcl-2 Family Antibody Sampler Kit recognizes only its specific target. The antibodies do not cross-react with other Bcl-2 family members. Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb detects endogenous levels of human Bcl-2 only when phosphorylated at Ser70. Phospho-Bcl-2 (Thr56) Antibody (Human Specific) detects endogenous levels of human Bcl-2 only when phosphorylated at Thr56.

Total Mcl-1, Bcl-xL, and Bcl-2 monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu210 of human Mcl-1, Asp61 of human Bcl-xL, and Gly47 of human Bcl-2. Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser70 of human Bcl-2. Phospho-Bcl-2 (Thr56) Antibody (Human Specific) is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr56 of human Bcl-2. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

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) that mediate protein interactions. The family can be separated into three groups based upon function and sequence homology: pro-survival members include Bcl-2, Bcl-xL, Mcl-1, A1 and Bcl-w; pro-apoptotic proteins include 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).

  1. Cory, S. et al. (2003) Oncogene 22, 8590-607.
  2. Antonsson, B. and Martinou, J.C. (2000) Exp Cell Res 256, 50-7.
  3. Sharpe, J.C. et al. (2004) Biochim Biophys Acta 1644, 107-13.
  4. Korsmeyer, S.J. et al. (1993) Semin Cancer Biol 4, 327-32.
  5. Bouillet, P. and Strasser, A. (2002) J Cell Sci 115, 1567-74.
  6. Maundrell, K. et al. (1997) J. Biol. Chem. 272, 25238-25242.
  7. Yamamoto, K. et al. (1999) Mol. Cell Biol. 19, 8469-8478.
  8. Ling, Y. H. et al. (1998) J. Biol. Chem. 273, 18984-18991.
  9. Huang, S.J. and Cidlowski, J.A. (2002) FASEB 16, 825-832.
  10. Deng, X. et al. (2001) J. Biol. Chem. 276, 23681-23688.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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