Cell Signaling Technology

Product Pathways - Apoptosis

Cleaved Caspase Antibody Sampler Kit #9929

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664 40 µl W IP IHC-P IHC-F IF-IC F H M R Mk (Dg) 17, 19 Rabbit IgG
Cleaved Caspase-6 (Asp162) Antibody #9761 40 µl W H M R 18 Rabbit
Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb #8438 40 µl W IF-IC F H M R (Mk) 18 Rabbit IgG
Cleaved Caspase-9 (Asp330) (D2D4) Rabbit mAb #7237 40 µl W IP H (Mk) 37 Rabbit IgG
Cleaved Caspase-9 (Asp315) Antibody (Human Specific) #9505 40 µl W IP H 35 Rabbit
Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625 40 µl W IP IHC-P IF-IC F H Mk 89 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl 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  Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Cleaved Caspase-3 (Asp175), Cleaved Caspase-6 (Asp162), Cleaved Caspase-7 (Asp198), Cleaved Caspase-9 (Asp315), Cleaved Caspase-9 (Asp330), and Cleaved PARP (Asp214) Antibodies detect endogenous levels of the large cleavage fragments of their respective targets. These antibodies will not cross-react with their respective full-length proteins.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with Staurosporine #9953 (1 μM, 3 hr), Jurkat cells, untreated or etoposide-treated (25 μM, overnight), and THP-1 cells, untreated or cycloheximide-treated (CHX, 10 μg/ml, overnight) followed by treatment with TNF-α #8902 (20 ng/ml, 4 hr), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625 (upper), or total PARP Antibody #9542 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and Jurkat cells, untreated or treated with Staurosporine #9953 (1 μM, 3 hr) or etoposide (25 μM, overnight), using Cleaved Caspase-9 (Asp330) (D2D4) Rabbit mAb #7237 (upper) or total Caspase-9 Antibody (Human Specific) #9502 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, C2C12, and H-4-II-E cells, untreated (-) or treated (+) with Staurosporine #9953 (1 μM, 3 hr), using Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb #8438 (upper) and Caspase-7 Antibody #9492 (lower).


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 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 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

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to amino-terminal residues adjacent to Asp175 of human caspase-3 or to residues surrounding Asp214 in human PARP, Asp330 of human caspase-9, or Asp198 of human caspase-7. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Asp315 of human caspase-9 or to the carboxy-terminal sequence of the large subunit (p18) of rat caspase-6. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

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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For Research Use Only. Not For Use In Diagnostic Procedures.

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