Product Pathways - Apoptosis / Autophagy
Cleaved Caspase-3 (Asp175) Antibody (Alexa Fluor® 488 Conjugate) #9669
PhosphoSitePlus® protein, site, and accession data: Casp3
| Applications | Reactivity | Sensitivity | Source |
|---|---|---|---|
| IF-IC F | H M R Mk B (Pg) | Endogenous | Rabbit |
Applications Key:
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
B=Bovine
Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 9669:
- Flow, Immunofluorescence
Specificity / Sensitivity
Cleaved Caspase-3 (Asp175) Antibody (Alexa Fluor® 488 Conjugate) detects endogenous levels of the large fragment (17/19 kDa) of activated caspase-3 resulting from cleavage adjacent to aspartic acid 175. The antibody does not recognize full length caspase-3 or other cleaved caspases. Alexa Fluor® 488 conjugates are excited at 494 nm and emit at 520 nm.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino-terminal residues adjacent to Asp175 of human caspase-3. Antibodies are purified by protein A and peptide affinity chromatography. The antibody was conjugated to Alexa Fluor® 488 under optimal conditions with an F/P ratio of 2-6.
Flow Cytometry
Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved Caspase-3 (Asp175) Antibody (Alexa Fluor® 488 Conjugate).
IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or staurosporine-treated (right), using Cleaved Caspase-3 (Asp175) Antibody (Alexa Fluor® 488 Conjugate) (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
Description
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Cleaved Caspase-3 (Asp175) Antibody #9661.
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
- Pei, X.Y. et al. (2011) Blood 118, 5189-200. Applications: IF-IC (In Cells)
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Companion Products
Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
