Cell Signaling Technology

Product Pathways - Apoptosis

Apoptosis Antibody Sampler Kit (Mouse Preferred) #9930

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Caspase-3 Antibody #9662 40 µl W IP IHC-P H M R Mk 17, 19, 35 Rabbit
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
Caspase-6 Antibody #9762 40 µl W H M R 15, 35 Rabbit
Cleaved PARP (Asp214) Antibody (Mouse Specific) #9544 40 µl W IF-IC M 89 Rabbit
Caspase-12 Antibody #2202 40 µl W M 42, 55 Rabbit
Caspase-9 Antibody (Mouse Specific) #9504 40 µl W M 37, 39, 49 Rabbit
Cleaved Caspase-9 (Asp353) Antibody (Mouse Specific) #9509 40 µl W IF-IC M 37 Rabbit
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

Each antibody in the kit detects endogenous levels of its respective protein. Cleaved Caspase-3 (Asp175) Antibody and Cleaved Caspase-6 (Asp162) Antibody detect the large fragment of their respective proteins. Caspase-3 Antibody and Caspase-9 Antibody (Mouse Specific) detect the full length and the large fragments. Cleaved caspase-9 (Asp353) detects the cleaved large 37 kDa fragment. Caspase-6 Antibody detects the full length fragment and the small subunit. Caspase-12 Antibody detects the full length fragment and its cleaved product. Cleaved PARP (Asp214) Antibody (Mouse Specific) detects the large fragment of mouse PARP produced by caspase cleavage.

Western Blotting

Western Blotting

Western blot analysis of extracts from L929 cells, untreated or Brefeldin A-treated (10 µg, 30 hours), using Caspase-12 Antibody #2202.

Western Blotting

Western Blotting

Western blot analysis of NIH/3T3 cells, untreated, staurosporine-treated (1 µM) or cytochrome C-treated (0.25 mg/ml), using Caspase-9 Antibody (Mouse Specific) #9504. (p39: caspase-9 cleaved at Asp368; p37: caspase-9 cleaved at Asp353.)

Western Blotting

Western Blotting

Western blot analysis of extracts from L929 cells, untreated or staurosporine-treated, using Cleaved Caspase-9 (Asp353) Antibody #9509 (upper) or Caspase-9 Antibody #9504 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or staurosporine-treated (1 µM), using Cleaved PARP (Asp214) Antibody (Mouse Specific) #9544.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat, L929 and C6 cells, untreated or cytochrome c-treated (0.25 mg/ml in vitro), using Caspase-3 Antibody #9662.

Western Blotting

Western Blotting

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


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 NIH/3T3 cells, untreated or staurosporine-treated (1 µM), and Jurkat cells, untreated or etoposide-treated (25 µM), using Caspase-6 Antibody #9762.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human tonsil, showing cytoplasmic localization, using Caspase-3 Antibody #9662.


IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical staining of wild-type P12 mouse cerebellum, using Caspase-9 Antibody (Mouse Specific) #9504. (Provided by Dr. Stella E. Tsirka, State University of New York, Dept. of Pharmacological Sciences.)

Description

The Apoptosis Antibody Sampler Kit (Mouse Specific) is designed for use with mouse samples and offers an economical means to evaluate the levels of active and inactive caspases. The kit contains enough primary and secondary antibodies to perform four Western blot experiments with each antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to amino-terminal residues of proteolytic cleavage sites of cleaved caspase-3 (Asp175), cleaved caspase-6 (rat Asp162), cleaved caspase-9 (Asp353), mouse-cleaved PARP, and residues surrounding amino acid 158 of mouse caspase-12. 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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