Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

Cleaved Caspase-9 (Asp330) Antibody (Human Specific) #9501

Applications Reactivity MW (kDa) Source
W IP H 17 large fragment. 37 large fragment + prodomain. Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

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.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to amino-terminal residues surrounding to Asp330 of human caspase-9. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

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).

Background

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. This complex processes procaspase-9 into a large active fragment (35 kDa or 17 kDa) and a small fragment (10 kDa) by self-cleavage at Asp315 (3-5). Cleaved caspase-9 further processes other caspase members, including caspase-3 and caspase-7, to initiate a caspase cascade, which leads to apoptosis (6-9). In addition to self-cleavage, procaspase-9 can also be cleaved in vivo by caspase-3 at Asp330. This process serves as positive feedback to amplify the apoptotic signal in the caspase activation pathway (3-5).

  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. Deveraux, Q. L. et al. (1998) EMBO J. 17, 2215-2223.
  7. Slee, E. A. et al. (1999) J. Cell Biol. 144, 281-292.
  8. Sun, X. et al. (1999) J. Biol. Chem. 274, 5053-5060.
  9. MacFarlane, M. et al. (1997) J. Cell Biol. 137, 469-479.

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