Product Pathways - Apoptosis
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.
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 treated with etoposide #2200 (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 #8953 (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 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
- 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.
For Research Use Only. Not For Use In Diagnostic Procedures.
