Cell Signaling Technology

Product Pathways - Apoptosis

Apoptosis Antibody Sampler Kit #9915

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
Caspase-3 Antibody #9662 40 µl W IP IHC-P H M R Mk 17, 19, 35 Rabbit
PARP Antibody #9542 40 µl W H M R Mk 89, 116 Rabbit
Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625 40 µl W IP IHC-P IF-IC F H Mk 89 Rabbit IgG
Caspase-9 Antibody (Human Specific) #9502 40 µl W F H 35, 37, 47 Rabbit
Cleaved Caspase-9 (Asp330) (D2D4) Rabbit mAb #7237 40 µl W IP H (Mk) 37 Rabbit IgG
Caspase-7 Antibody #9492 40 µl W H M R Mk 20, 35 Rabbit
Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb #8438 40 µl W IF-IC F H M R (Mk) 18 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

Each antibody in the Apoptosis Antibody Sampler Kit detects endogenous levels of its respective target. Cleaved Caspase-3 (Asp175), Cleaved Caspase-7 (Asp198), Cleaved Caspase-9 (Asp330), and Cleaved PARP (Asp214) Antibodies detect only the large fragments of their respective targets. Caspase-3, Caspase-7, Caspase-9 and PARP Antibodies detect both the full length and the large cleaved fragments of their respective targets.

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 from Jurkat cells, untreated or etoposide-treated (25 µM), and HeLa cells, untreated or staurosporine-treated (1 µM), using Caspase-7 Antibody #9492.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated, staurosporine-treated (1 µM) or cytochrome c-treated (0.25 mg/ml), and Jurkat cells, untreated or cytochrome c-treated, using Caspase-9 Antibody (Human Specific) #9502.

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 PARP Antibody #9542.


Western Blotting

Western Blotting

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

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or treated with cytochrome c, showing full length and cleaved caspase-3 and cleaved caspase-3 (19 and 17 kDa) fragments, 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 or etoposide as indicated, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human tonsil, showing nuclear staining, using PARP Antibody #9542.

IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse embryo, using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb #9664 in the presence of control peptide (left) or Cleaved Caspase-3 (Asp175) Blocking Peptide (#1050) (right).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved Caspase-7 (Asp198) Antibody #9491 compared to a nonspecific negative control antibody (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide treated (green), using Caspase-7 Antibody #9492 compared to a nonspecific negative control antibody (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide treated (green), using Cleaved Caspase-9 (Asp330) Antibody (Human Specific) #9501 compared to a nonspecific negative control antibody (red).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Caspase-9 Antibody (Human Specific) #9502 compared to a nonspecific negative control antibody (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved Caspase-3(Asp175) (5A1) Rabbit mAb #9664 compared to a nonspecific negative control antibody (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide treated (green), using Cleaved Caspase-3 (Asp175) Antibody #9661 compared to a nonspecific negative control antibody (red).


Description

The Apoptosis Antibody Sampler Kit provides an economical means to evaluate the levels of inactive and active caspases. The kit contains enough primary and secondary antibodies to perform four Western blot experiments with each antibody.

Source / Purification

Monoclonal and polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding the proteolytic cleavage sites of human caspase-3, -7 and -9 and human PARP. 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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