Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

Cleaved Caspase-9 (Asp315) Antibody (Human Specific) #9505

Applications Reactivity Sensitivity MW (kDa) Source
W IP H Endogenous 35 Rabbit

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

Protocols

Specificity / Sensitivity

Cleaved Caspase-9 (Asp315) Antibody (Human Specific) detects endogenous levels of the 35 kDa large fragment of caspase-9 following cleavage at aspartic acid 315. The antibody does not react with full length caspase-9 or any other cleaved caspases.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp315 of human caspase-9. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or cytochrome c-treated (0.25 mg/ml), and HeLa cells, untreated, staurosporine-treated (1 µM), or cytochrome c-treated (0.25 mg/ml), using Cleaved Caspase-9 (Asp315) Antibody (Human Specific) (upper) or Caspase-9 Antibody (Human Specific) #9502 (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. 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.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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