Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

Cleaved Caspase-9 (Asp353) Antibody (Mouse Specific) #9509

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC M Endogenous 37 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  M=Mouse
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

Cleaved Caspase-9 (Asp353) Antibody (Mouse Specific) detects endogenous levels of the 37kDa subunit of mouse caspase-9 only after after cleavage at aspartic acid 353. It does not cross-react with full length caspase-9 or with other caspases at endogenous levels.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp353 of mouse caspase-9. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from L929 cells, untreated or staurosporine-treated, using Cleaved Caspase-9 (Asp353) Antibody (upper) or Caspase-9 Antibody #9504 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of NIH/3T3 cells, staurosporine-treated (left) or untreated (right), using Cleaved Caspase-9 (Asp353) Antibody (Mouse Specific) (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

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.

Application References

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Companion Products

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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