Product Pathways - Apoptosis / Autophagy

Cleaved Caspase Antibody Sampler Kit #9929

• pathway
• more info
• application references
• datasheet PDF
• MSDS PDF
• protocols

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

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 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 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 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 blot analysis of extracts from HeLa, NIH/3T3 and C6 cells, untreated, staurosporine-treated (1 µM in vivo) or cytochrome c-treated (0.25 mg/ml in vitro), using Cleaved Caspase-3 (Asp175) Antibody #9661.

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.

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human tonsil, showing perinuclear staining of select epithelial cells, using Cleaved Caspase-3 (Asp175) Antibody #9661.

IHC-FL (floating)

Immunostaining of cerebellar neuron culture from newborn rat, untreated or tert-Butyl-hydroperoxide (tBH)-treated (40 µM, 30 minute treatement, 3 hour recovery), showing tBH-induced neuronal apoptosis and cleaved caspase-7 co-localization with apoptotic neurons, using Cleaved Caspase-7 (Asp198) Antibody #9491 (green) and propidium iodide (red).

IHC-FL (floating)

Confocal micrograph of newborn rat brain cortex (Cx) and striatum (Str), using Cleaved Caspase-3 (Asp175) Antibody #9661 (green) and propidium iodide (red). (Provided by Dr. Bingren Hu, University of Miami School of Medicine, Florida.)

IC-ABC

Immunocytochemical staining of HeLa cells, untreated or staurosporine-treated (1 µM), using Cleaved Caspase-9 (Asp330) Antibody (Human Specific) #9501.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic peptides (KLH-coupled) 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, alpha-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-alpha, 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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

• 2202 Caspase-12 Antibody
• 9492 Caspase-7 Antibody
• 9502 Caspase-9 Antibody (Human Specific)
• 9504 Caspase-9 Antibody (Mouse Specific)
• 9506 Caspase-9 Antibody (Rat Specific)
• 9507 Cleaved Caspase-9 (Asp353) Antibody (Rat Specific)
• 9662 Caspase-3 Antibody
• 9752 Caspase-10 Antibody
• 9762 Caspase-6 Antibody
• 9852 Chaps Cell Extract Buffer (10X)
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide