Product Pathways - Apoptosis

Apoptosis Marker: SignalStain^® Cleaved Caspase-3 (Asp175) IHC Detection Kit #8120

Kit Includes	Cap Color
Cleaved Caspase-3 Blocking Peptide	Dark Blue
Peroxide Quench	Orange
Blocking Solution	Blue
Prediluted Negative Control	Brown
Prediluted Cleaved Caspase-3 (D175) Ab #9661	Purple
Biotinylated Secondary Ab	Green
Reagent A	Gray
Reagent B	Gray
NovaRED Substrate 1 (TM)	Red
NovaRED Substrate 2 (TM)	Red
NovaRED Substrate 3 (TM)	Red
NovaRED Substrate 4 (TM)	Red
Mixing Bottle	Yellow

NovaRED™ is a trademark of Vector Laboratories.

Description

CST’s Apoptosis Marker: SignalStain® Cleaved Caspase-3 (Asp175) IHC Detection Kit is a "ready to use" system designed to detect the activation of caspase-3 in human tissue and cell preps by immunohistochemistry. The kit utilizes the ABC immunoperoxidase method to detect endogenous levels of caspase-3 protein. Prediluted Cleaved Caspase-3 (Asp175) Antibody is bound by a biotinylated secondary antibody. Avidin DH and biotinylated horseradish peroxidase are complexed by mixing defined amounts prior to use, and the mixture subsequently binds the secondary antibody. The macromolecular complex is localized by incubation with NovaRED™ enzyme substrate.The prediluted primary antibody, along with the ABC system, allows the user to examine caspase-3 localization consistently and offers the highest sensitivity with the lowest background.

Species Cross-Reactivity

H M (R)

Reactivity Key: H=Human M=Mouse R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Apoptosis Marker: SignalStain® Cleaved Caspase-3 (Asp175) IHC Detection Kit 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. This kit was developed for and is recommended for immunohistochemistry only.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino-terminal residues surrounding to Asp175 in human caspase-3. Antibodies are purified by protein A and peptide affinity chromatography.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse embryo, using Apoptosis Marker: SignalStain^® Cleaved Caspase-3 (Asp175) IHC Detection Kit (left) or matched negative control (right).

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).

Application References

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. , 449-454. Applications: IHC-P (paraffin)
DiCunto, F. et al. (2000) Defective neurogenesis in citron kinase knockout mice by altered cytokinesis and massive apoptosis. Neuron 28, 115-127. Applications: Flow Cytometry IHC-P (paraffin)
Olney, J. W. et al. (2002) Induced apoptotic neurodegeneration in the developing C57BL/6 mouse brain. Brain Res. Dev. Brain Res. 133 (2), 115-126. Applications: IHC-P (paraffin)

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.