Cleaved Caspase-9 (Asp330) Ant

9501:

Immunoprecipitation , Western Blotting

Cleaved Caspase-9 (Asp330) Antibody (Human Specific) detects endogenous levels of the large fragment (37 kDa with prodomain/17 kDa) of caspase-9 following cleavage at aspartic acid 330. The antibody does not recognize the uncleaved procaspase-9.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino terminus residues surrounding to Asp330 of human caspase-9. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of HeLa cells, untreated or staurosporine-treated (1 µM), and Jurkat cells, untreated or etoposide-treated, using Cleaved Caspase-9 (Asp330) Antibody (Human Specific) (upper) or Caspase-9 Antibody #9502 (Human Specific) (lower).

Caspase-9 (ICE-LAP6, Mch6) is an important member of the cysteine aspartic acid protease (caspase) family (1,2). Upon apoptotic stimulation, cytochrome c released from mitochondria associates with the 47 kDa procaspase-9/Apaf 1. Apaf-1 mediated activation of caspase-9 involves instrinsic proteolytic processing resulting in cleavage at Asp315 and producing a p35 subunit. Another cleavage occurs at Asp330 producing a p37 subunit that can serve to amplify the apoptotic response (3-6). Cleaved caspase-9 further processes other caspase members, including caspase-3 and caspase-7, to initiate a caspase cascade, which leads to apoptosis (7-10).

1. Duan, H. et al. (1996) J. Biol. Chem. 271, 16720-16724.
2. Srinivasula, S. M. et al. (1996) J. Biol. Chem. 271, 27099-27106.
3. Liu, X. et al. (1996) Cell 86, 147-157.
4. Li, P. et al. (1997) Cell 91, 479-489.
5. Zou, H. et al. (1999) J. Biol. Chem. 274, 11549-11556.
6. Srinivasula, S.M. et al. (1998) Mol Cell 1, 949-57.
7. Deveraux, Q. L. et al. (1998) EMBO J. 17, 2215-2223.
8. Slee, E. A. et al. (1999) J. Cell Biol. 144, 281-292.
9. Sun, X. et al. (1999) J. Biol. Chem. 274, 5053-5060.
10. MacFarlane, M. et al. (1997) J. Cell Biol. 137, 469-479.

• Li, J. et al. (2000) Cyclic nucleotides suppress tumor necrosis factor alpha-mediated apoptosis by inhibiting caspase activation and cytochrome c release in primary hepatocytes via a mechanism independent of Akt activation. J. Biol. Chem. 275, 13026-13034. Applications: Western Blotting

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• 9502 Caspase-9 Antibody (Human Specific)
• 9504 Caspase-9 Antibody (Mouse Specific)
• 9505 Cleaved Caspase-9 (Asp315) Antibody (Human Specific)
• 9506 Caspase-9 Antibody (Rat Specific)
• 9507 Cleaved Caspase-9 (Asp353) Antibody (Rat Specific)
• 9852 Chaps Cell Extract Buffer (10X)
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

For Research Use Only. Not For Use In Diagnostic Procedures.