Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

Cleaved Caspase Antibody Sampler Kit #9929

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb # 9664 40 microliters W IP IHC-P IHC-F IF-IC F H M R Mk 17, 19 Rabbit IgG
Cleaved Caspase-6 (Asp162) Antibody # 9761 40 microliters W H M R 18 Rabbit
Cleaved Caspase-7 (Asp198) Antibody # 9491 40 microliters W IP H M R Mk 20 Rabbit
Cleaved Caspase-9 (Asp330) Antibody (Human Specific) # 9501 40 microliters W IP H Mk 17 large fragment. 37 large fragment + prodomain. Rabbit
Cleaved Caspase-9 (Asp315) Antibody (Human Specific) # 9505 40 microliters W IP H 35 Rabbit
Cleaved PARP (Asp214) Antibody (Human Specific) # 9541 40 microliters W IHC-P IF-IC F H 89 Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 microliters Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey

Specificity / Sensitivity

Cleaved Caspase-3 (Asp175), Cleaved Caspase-6 (Asp162), Cleaved Caspase-7 (Asp198) and Cleaved Caspase-9 (Asp315) Antibodies detect the large fragments of their respective cleaved proteins. Cleaved PARP (Asp214) Antibody detects the cleaved 89 kDa large fragment of PARP. Cleaved Caspase-9 (Asp330) Antibody detects the 37 kDa cleaved large fragment + prodomain and the 17 kDa large fragment. These antibodies will not cross-react with their respective full-length proteins.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat and 293 cells, untreated, cytochrome c-treated (0.25 mg/ml, 1 hour) or etoposide-treated (25 µM, 5 hours), using Cleaved Caspase-7 (Asp198) Antibody #9491.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and Jurkat cells, untreated, staurosporine-treated (1 µM) or etoposide-treated, using Cleaved Caspase-9 (Asp330) Antibody (Human Specific) #9501.

Western Blotting

Western Blotting

Western blot analysis of extracts fron HeLa and Jurkat cells, untreated, staurosporine-treated (1 µM) or cytochrome c-treated (0.25 mg/ml), using Cleaved Caspase-9 (Asp315) Antibody (Human Specific) #9505.


Western Blotting

Western Blotting

Western blot analysis of Jurkat cells, untreated or etoposide-treated (25 µM), using Cleaved PARP (Asp214) Antibody (Human Specific) #9541.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated (-) or etoposide-treated (+), and NIH/3T3 and C6 cells, untreated or staurosporine-treated, using Cleaved Caspase-6 (Asp162) Antibody #9761.

Western Blotting

Western Blotting

Western blot analysis of extracts from C6 (rat), NIH/3T3 (mouse), and Jurkat (human) cells, untreated or treated with staurosporine (1uM, 3hrs) or etoposide (25uM, 5hrs) as indicated, using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb #9664.


Description

The Cleaved Caspase Antibody Sampler Kit provides an economical means to evaluate the activation status of cleaved caspases by detecting their cleaved forms. The kit contains enough primary and secondary antibodies to perform four Western blot experiments with each primary antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the amino-terminal residues adjacent to the proteolytic cleavage sites of human PARP, caspase-3, -7, -9 and rat caspase-6. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is supplied in HEPES buffer with 50% glycerol and less than 0.02% sodium azide.

Background

Apoptosis is a regulated physiological process leading to cell death. Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Initiator caspases (including 8, 9, 10 and 12) are closely coupled to proapoptotic signals. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6 and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF and lamin A, and induce apoptosis. Cytochrome c released from mitochondria is coupled to the activation of caspase-9, a key initiator caspase (1). Proapoptotic stimuli include the FasL, TNF-α, DNA damage and ER stress. Fas and TNFR activate caspases 8 and 10 (2), DNA damage leads to the activation of caspase-9 and ER stress leads to the calcium-mediated activation of caspase-12 (3). The inhibitor of apoptosis protein (IAP) family includes XIAP and survivin and functions by binding and inhibiting several caspases (4,5). Smac/Diablo, a mitochondrial protein, is released into the cytosol upon mitochondrial stress and competes with caspases for binding of IAPs. The interaction of Smac/Diablo with IAPs relieves the inhibitory effects of the IAPs on caspases (6).

  1. Baker, S.J. and Reddy, E.P. (1998) Oncogene 17, 3261-3270.
  2. Budihardjo, I. et al. (1999) Annu. Rev. Cell Dev. Biol. 15, 269-290.
  3. Nakagawa, T. et al. (2000) Nature 403, 98-103.
  4. Deveraux, Q. L. et al. (1998) EMBO J. 17, 2215-2223.
  5. Li, F. et al. (1998) Nature 396, 580-584.
  6. Du, C. et al. (2000) Cell 102, 33-42.

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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