Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

Cleaved Caspase-9 (Asp353) Antibody (Rat Specific) #9507

Applications Reactivity MW (kDa) Source
W IC R 17, 38 Rabbit

Applications Key:  W=Western Blotting  IC=Immunocytochemistry
Reactivity Key:  R=Rat
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 (Asp353) Antibody (Rat Specific) detects endogenous levels of the large fragment (17 kDa or 38 kDa with prodomain) of caspase-9 resulting from cleavage at aspartic acid 353. The antibody does not recognize full length caspase-9 or any other cleaved caspases.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to amino-terminal residues surrounding Asp353 of rat caspase-9. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of C6 cells, untreated or cytochrome c-treated (0.25 mg/ml), and PC12 cells, untreated or staurosporine-treated, using Cleaved Caspase-9 (Asp353) Antibody (Rat Specific).

IHC-FL (floating)

IHC-FL (floating)

Confocal immunofluorescent images of 7-day postnatal rat pup cortex regions, control (left) and hypoxia/ischemia (2 hours followed by 48 hours of recovery, right), using Cleaved Caspase-9 (Asp353) Antibody (Rat Specific) (green) and propidium iodide (red). (Provided by Dr. Bingren Hu, University of Miami School of Medicine, Florida.)

IHC-FL (floating)

IHC-FL (floating)

Confocal immunofluorescent images of 7-day postnatal rat pup cortex regions, control and hypoxia/ischemia (2 hours followed by 48 hours of recovery), using Cleaved Caspase-9 (Asp353) Antibody (Rat Specific) (green) and propidium iodide (red). (A) 10X, (B) 40X, (C) 100X and (D) 100X plus 2.5 zoom. (Provided by Dr. Bingren Hu, University of Miami School of Medicine, Florida.)


IC-ABC

IC-ABC

Immunocytochemical staining of C6 cells, untreated (A, B) or staurosporine-treated (C, D), using Cleaved Caspase-9 (Asp353) Antibody (Rat Specific) (A, C) or the same antibody preincubated with competing peptide (B, D).

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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