Product Pathways - Apoptosis
Cleaved Caspase-3 (Asp175) Antibody #9661
PhosphoSitePlus® protein, site, and accession data: Casp3
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IP IHC-P IHC-F IF-IC F | H M R Mk (B) (Dg) (Pg) | Endogenous | 17, 19 | Rabbit |
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 B=Bovine Dg=Dog Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Cleaved Caspase-3 (Asp175) Antibody detects endogenous levels of the large fragment (17/19 kDa) of activated caspase-3 resulting from cleavage adjacent to Asp175. This antibody does not recognize full length caspase-3 or other cleaved caspases.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino-terminal residues adjacent to (Asp175) in human caspase-3.
Western Blotting
Western blot analysis of extracts from HeLa, NIH/3T3 and C6 cells untreated, staurosporine-treated (3hrs, 1 µM in vivo) or cytochrome c-treated (1hr, 0.25 mg/ml in vitro), using Caspase-3 Antibody #9662 (upper) or Cleaved Caspase-3 (Asp175) Antibody (lower).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human tonsil, showing cytoplasmic and perinuclear localization in apoptotic cells (low and high magnification), using Cleaved Caspase-3 (Asp175) Antibody.
IHC-P (paraffin)
Immunohistochemical analysis using Cleaved caspase-3 (Asp175) antibody on SignalSlide™ Cleaved Caspase-3 IHC controls #8104 (paraffin-embedded Jurkat cells, untreated (left) or etoposide-treated (right)).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded mouse embryo, using Cleaved Caspase-3 (Asp175) Antibody preincubated with control peptide (left) or Cleaved Caspase-3 (Asp175) Blocking Peptide #1050 (right).
IHC-F (frozen)
Immunohistochemical analysis of frozen H1650 xenograft section, using Cleaved Caspase-3 (Asp175) Antibody.
Flow Cytometry
Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved Caspase-3 (Asp175) Antibody compared to a nonspecific negative control antibody (red).
IF-IC
Confocal immunofluorescent images of HT-29 cells, untreated (left) or Staurosporine #9953 treated (right), labeled with Cleaved Caspase-3 (Asp175) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
Background
Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins, such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (1). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments. Cleavage of caspase-3 requires the aspartic acid residue at the P1 position (2).
- Fernandes-Alnemri, T. et al. (1994) J. Biol. Chem. 269, 30761-30764.
- Nicholson, D. W. et al. (1995) Nature 376, 37-43.
Application References
- Di Cunto, F. et al. (2000) Neuron 28, 115-27. Applications: Flow Cytometry IHC-P (paraffin)
- Cheong, J.W. et al. (2003) Clin Cancer Res 9, 5018-27. Applications: Western Blotting
- Garnier, P. et al. (2003) J Neurosci 23, 7967-73. Applications: Western Blotting
- Goodyear, C.S. et al. (2004) J Immunol 172, 2870-7. Applications: Flow Cytometry
- Gown, A.M. and Willingham, M.C. (2002) J Histochem Cytochem 50, 449-54. Applications: IHC-P (paraffin)
- Olney, J.W. et al. (2002) Brain Res Dev Brain Res 133, 115-26. Applications: IHC-P (paraffin)
- Kaiser, C.L. et al. (2008) Hear Res 240, 1-11. Applications: IHC-P (paraffin)
- Patel, A.V. and Krimm, R.F. (2010) Dev Biol 340, 419-29. Applications: IHC-P (paraffin)
- Faber, A. et al. (2011) Cancer Discov 1, 352-365. Applications: IHC-P (paraffin)
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
- 9664 Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb
- 9662 Caspase-3 Antibody
- 9665 Caspase-3 (8G10) Rabbit mAb
- 9668 Caspase-3 (3G2) Mouse mAb
- 1050 Cleaved Caspase-3 (Asp175) Blocking Peptide
- 9852 Chaps Cell Extract Buffer (10X)
- 8104 SignalSlide® Cleaved Caspase-3 (Asp175) IHC Controls
- 8112 SignalStain® Antibody Diluent
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 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
- 8114 SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
- 9579 Cleaved Caspase-3 (Asp175) (D3E9) Rabbit mAb
For Research Use Only. Not For Use In Diagnostic Procedures.
