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 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 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 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 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 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 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 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 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 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)
Immunohistochemical staining of paraffin-embedded human tonsil, showing nuclear staining, using PARP Antibody #9542.
IHC-P (paraffin)
Immunohistochemical staining of paraffin-embedded human tonsil, showing cytoplasmic localization of caspase-3, using Caspase-3 Antibody #9662.
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 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 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 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 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 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 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).
- Baker, S.J. and Reddy, E.P. (1998) Oncogene 17, 3261-3270.
- Budihardjo, I. et al. (1999) Annu. Rev. Cell Dev. Biol. 15, 269-290.
- Nakagawa, T. et al. (2000) Nature 403, 98-103.
- Deveraux, Q. L. et al. (1998) EMBO J. 17, 2215-2223.
- Li, F. et al. (1998) Nature 396, 580-584.
- 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!
Protocols
- 5625:
- Flow, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting
- 7237:
- Immunoprecipitation, Western Blotting
- 8438:
- Flow, Immunofluorescence, Western Blotting
- 9492:
- Western Blotting
- 9502:
- Flow, Western Blotting
- 9542:
- Western Blotting
- 9662:
- IHC / Paraffin, Immunoprecipitation, Western Blotting
- 9664:
- Flow, IHC / Frozen, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting
Companion Products
- 9504 Caspase-9 Antibody (Mouse Specific)
- 9505 Cleaved Caspase-9 (Asp315) Antibody (Human Specific)
- 9507 Cleaved Caspase-9 (Asp353) Antibody (Rat Specific)
- 9544 Cleaved PARP (Asp214) Antibody (Mouse Specific)
- 9545 Cleaved PARP (Asp214) Antibody (Rat Specific)
- 9546 Cleaved PARP (Asp214) (19F4) Mouse mAb (Human Specific)
- 9852 Chaps Cell Extract Buffer (10X)
- 9803 Cell Lysis 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
For Research Use Only. Not For Use In Diagnostic Procedures.
