Product Pathways - Apoptosis
Cleaved Caspase-9 (Asp315) Antibody (Human Specific) #9505
PhosphoSitePlus® protein, site, and accession data: Casp9
| 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 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).
- Duan, H. et al. (1996) J. Biol. Chem. 271, 16720-16724.
- Srinivasula, S. M. et al. (1996) J. Biol. Chem. 271, 27099-27106.
- Liu, X. et al. (1996) Cell 86, 147-157.
- Li, P. et al. (1997) Cell 91, 479-489.
- Zou, H. et al. (1999) J. Biol. Chem. 274, 11549-11556.
- Srinivasula, S.M. et al. (1998) Mol Cell 1, 949-57.
- Deveraux, Q. L. et al. (1998) EMBO J. 17, 2215-2223.
- Slee, E. A. et al. (1999) J. Cell Biol. 144, 281-292.
- Sun, X. et al. (1999) J. Biol. Chem. 274, 5053-5060.
- MacFarlane, M. et al. (1997) J. Cell Biol. 137, 469-479.
Application References
- Mitchell, K.O. et al. (2000) Cancer Res 60, 6318-25. Applications: Western Blotting
- Cheong, J.W. et al. (2003) Clin Cancer Res 9, 5018-27. Applications: Western Blotting
- Kaiser, C.L. et al. (2008) Hear Res 240, 1-11. Applications: IHC-P (paraffin)
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Companion Products
- 9501 Cleaved Caspase-9 (Asp330) Antibody (Human Specific)
- 9502 Caspase-9 Antibody (Human Specific)
- 9504 Caspase-9 Antibody (Mouse 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.