Product Pathways - Apoptosis / Autophagy
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IHC-P IHC-F IF-IC F | H M R | Endogenous | 17, 19 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IHC-F=Immunohistochemistry (Frozen)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb 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.
Source / Purification
Monoclonal antibody is produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to amino-terminal residues adjacent to Asp175 of human caspase-3.
Western Blotting
Western blot analysis of extracts from C6 (rat), NIH/3T3 (mouse), and Jurkat (human) cells, untreated or treated with staurosporine or etoposide as indicated, using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb.
IP
Immunoprecipitation of extracts from Jurkat cells, untreated or etoposide-treated, using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb. Western blot was performed using the same antibody.
IHC-P (paraffin)
Immunofluorescent analysis of paraffin-embedded mouse embryo, using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb (red) and DAPI (blue).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded mouse embryo, using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb in the presence of control peptide (left) or Cleaved Caspase-3 (Asp175) Blocking Peptide (#1050) (right).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded Jurkat cells, untreated (left) or etoposide-treated (right), using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical staining of paraffin-embedded mouse embryo, showing cytoplasmic localization in apoptotic cells, using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb.
IHC-F (frozen)
Immunohistochemical analysis of frozen H1650 xenograft, using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb.
IHC-FL (floating)
Confocal micrograph of newborn rat brain cortex, using Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb (green) and propidium iodide (red). (Provided by Dr. Bingren Hu, University of Miami School of Medicine, Florida.)
Flow Cytometry
Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved Caspase-3(Asp175) (5A1) Rabbit mAb compared to a nonspecific negative control antibody (red).
IF-IC
Confocal immunofluorescent images of HT-29 cells, untreated (left) or Staurosporine #9953 treated (right) labeled with Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
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
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- 1050 Cleaved Caspase-3 (Asp175) Blocking Peptide
- 9547 Cleaved PARP (Asp214) Antibody (Human Specific) (Fluorescein Conjugate)
- 8104 SignalSlide™ Cleaved Caspase-3 (Asp175) IHC Controls
Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.
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.
