Product Pathways - Apoptosis / Autophagy
Apoptosis Marker: SignalStain® Cleaved Caspase-3 (Asp175) IHC Detection Kit #8120
| Kit Includes | Cap Color |
|---|---|
| Cleaved Caspase-3 Blocking Peptide | White |
| Peroxide Quench | Orange |
| Blocking Solution | Blue |
| Prediluted Negative Control | Brown |
| Prediluted Cleaved Caspase-3 (Asp175) Antibody #9661 | Purple |
| Biotinylated Secondary Antibody | 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 | Red |
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.
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 rabbits with a synthetic peptide (KLH coupled) 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 aspartic acid 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
- 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. This article references the use of Apoptosis Marker: SignalStain® Cleaved Caspase-3 (Asp175) IHC Detection Kit in the following 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. This article references the use of Apoptosis Marker: SignalStain® Cleaved Caspase-3 (Asp175) IHC Detection Kit in the following 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. This article references the use of Apoptosis Marker: SignalStain® Cleaved Caspase-3 (Asp175) IHC Detection Kit in the following 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
- 1050 Cleaved Caspase-3 (Asp175) Blocking Peptide
- 9661 Cleaved Caspase-3 (Asp175) Antibody
- 8104 SignalSlide™ Cleaved Caspase-3 (Asp175) IHC Controls
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
