Product Pathways - Apoptosis / Autophagy
Cleaved Caspase-3 (Asp175) Antibody (Alexa Fluor® 488 Conjugate) #9669
| Applications | Reactivity | Sensitivity | Source |
|---|---|---|---|
| IF-IC F | H (M) (R) | Endogenous | Rabbit |
Applications Key:
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
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 (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 (KLH-coupled) 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
Immunofluorescent analysis of HeLa cells, untreated (A) or staurosporine-treated (B), using Cleaved Caspase-3 (Asp175) Antibody (Alexa Fluor® 488 Conjugate) (green) and DAPI (blue).
Description
Cell Signaling Technology Antibody conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct Flow Cytometric analysis of human cells.The unconjugated antibody, #9661, reacts with human, mouse, and rat cleaved caspase-3. CST expects that Cleaved Caspase 3 (Alexa Fluor® 488 Conjugate) will also recognize cleaved caspase-3 in these species.
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
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
- 9662 Caspase-3 Antibody
Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
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.
