Product Pathways - Apoptosis / Autophagy

Cleaved Caspase-3 (Asp175) Antibody #9661

Applications	Reactivity	MW (kDa)	Source
W IHC-P IHC-F IF-IC IC F	H M R B (Pg)	17, 19	Rabbit

Applications Key: W=Western Blotting 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 Pg=Pig B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

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 rabbits with a synthetic peptide (KLH-coupled) corresponding to amino-terminal residues adjacent to (Asp175) in human caspase-3. Antibodies are purified by protein A and peptide affinity chromatography.

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 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 of paraffin-embedded Jurkat cells, untreated (left) or etoposide treated (right), using Cleaved Caspase-3 (Asp175) Antibody.

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.

IHC-FL (floating)

Confocal micrograph of newborn rat brain tissue, showing control and transient cerebral ischemia, using Cleaved Caspase-3 (Asp175) Antibody (green) and propidium iodide (red). (Provided by Dr. Bingren Hu, University of Miami School of Medicine, Florida.)

IHC-FL (floating)

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

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).

Flow Cytometry

Flow cytometric analysis of Pam212 cells, untreated or sulindac sulfone-treated (to induce apoptosis), using Cleaved Caspase-3 (Asp175) Antibody. Results were similar to those obtained by analyzing DNA content. (Provided by Dan Rosson, Ph.D., Lankenau Institute of Medical Research, Wynnewood, PA.)

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 aspartic acid at the P1 position (2).

Application References

DiCunto, F. et al. (2000) Defective neurogenesis in citron kinase knockout mice by altered cytokinesis and massive apoptosis. Neuron 28, 115-127. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: Flow Cytometry IHC-P (paraffin)
Reis, T. and Edgar, B.A. (2004) Negative Regulation of dE2F1 by Cyclin-Dependent Kinases Controls Cell Cycle Timing. Cell 117 (2), 253-264. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: Fluorescent Detection
Tinsley, K. W. et al. (2003) Sepsis induces apoptosis and profound depletion of splenic interdigitating and follicular dendritic cells. J. Immunology 171 (2), 909-14. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: IHC-FL (floating/frozen)
Cheong, J. W. et al. (2003) Induction of apoptosis by apicidin, a histone deacetylase inhibitor, via the activation of mitochondria-dependent caspase cascades in human Bcr-Abl-positive leukemia cells. Clin. Cancer Res. 9 (13), 5018-27. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: Western Blotting
Sanchez-Gomez, M. V. et al. (2003) Caspase-dependent and caspase-independent oligodendrocytes death mediated by AMPA and kainate receptors. J. Neurosci. 23 (29), 9519-28. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: IC-IF
Garnier, P. et al. (2003) Ischemic preconditioning by caspase cleavage of poly)ADP-ribose) polymerase-1. J. Neuroscience 23 (22), 7967-73. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: Western Blotting
Goodyear, C. S. et al. (2004) In vivo VL-targeted activation-induced apoptotic supraclonal deletion by a microbial B cell toxin. J. Immunology 172 (5), 2870-7. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: Flow Cytometry
Saito, A. et al. (2004) Neuroprotective role of a proline-rich Akt substrate in apoptotic neuronal cell death after stroke: relationships with nerve growth factor. J. Neurosci. 24, 1584-1593. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: IC-IF
Gown, A.M. and Willingham, M.C. (2002) Improved detection of apoptotic cells in archival paraffin sections: immunohistochemistry using antibodies to cleaved caspase 3. J. Histochem. Cytochem. 50 (4), 449-454. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: IHC-P (paraffin)
Olney, J. W. et al. (2002) Ethanol-induced apoptotic neurodegeneration in the developing C57BL/6 mouse brain. Brain Res. Dev. Brain Res. 133 (2), 115-126. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: IHC-P (paraffin)
Wang, J. et al. (2002) Increased in vivo apoptosis in cells lacking mitochondrial DNA gene expression. Proc. Nat. Acad. Sci. USA 98 (7), 4038-4043. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: IHC-FL (floating/frozen)
Hu, B. R. et al. (2000) Involvement of caspase-3 in cell death after hypoxia-ischemia declines during brain maturation. J. Cereb. Blood Flow Metab. 20 (9), 1294-1300. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: IHC-FL (floating/frozen) Western Blotting
Decaudin, D. et al. (2002) Peripheral benzodiazepine receptor ligands reverse apoptosis resistance of cancer in vitro and in vivo. Cancer Res. 62, 1388-1393. This article references the use of Cleaved Caspase-3 (Asp175) Antibody in the following applications: IC-IF

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