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Western blot analysis of extracts from C6 (rat), NIH/3T3 (mouse), and Jurkat (human) cells, untreated or treated with staurosporine #9953 (1uM, 3hrs) or etoposide #2200 (25uM, 5hrs) as indicated, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.

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Western blot analysis of extracts from Jurkat cells, untreated or etoposide-treated (25uM, 5hrs), and NIH/3T3 cells, untreated or staurosporine-treated (1uM, 3hrs), using Caspase-3 Antibody.

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After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Immunoprecipitation of extracts from Jurkat cells, untreated or etoposide-treated (25uM, 5hrs), using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb. Western blot was performed using the same antibody.

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Immunohistochemical staining of paraffin-embedded human tonsil, showing cytoplasmic localization, using Caspase-3 Antibody.

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Immunohistochemical analysis of paraffin-embedded mouse embryo, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb in the presence of control peptide (left) or Cleaved Caspase-3 (Asp175) Blocking Peptide (#1050) (right).

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Immunohistochemical analysis using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb on SignalSlide® Cleaved Caspase-3 IHC Controls #8104 (paraffin-embedded Jurkat cells, untreated (left) or etoposide-treated (right)).

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Immunohistochemical staining of paraffin-embedded mouse embryo, showing cytoplasmic localization in apoptotic cells, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.

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Immunohistochemical analysis of frozen H1650 xenograft, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.

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Confocal immunofluorescent images of HT-29 cells, untreated (left) or Staurosporine #9953 treated (right) labeled with Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Image
Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb 9664 100 µl
H M R Mk 17, 19 Rabbit IgG
Caspase-3 Antibody 9662 100 µl
H M R Mk 17, 19, 35 Rabbit 
Caspase-3 Control Cell Extracts 9663 100 µl
 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
All Goat 
Anti-biotin, HRP-linked Antibody 7075 100 µl
Goat 
Biotinylated Protein Ladder Detection Pack 7727 100 µl
 
20X LumiGLO® Reagent and 20X Peroxide 7003 5 ml each
 

Product Description

Apoptosis Marker: Cleaved Caspase-3 (Asp175) Western Detection Kit offers an efficient way of detecting caspase-3 processing and activation by Western blotting. The kit contains enough primary and secondary antibodies to perform 10 Western mini blots, as well as a set of pre-stained and biotinylated markers, cell lysates and LumiGLO® reagent.


Specificity / Sensitivity

Cleaved Caspase-3 (Asp175) Antibody 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. Caspase-3 Antibody detects endogenous levels of full length caspase-3 (35 kDa) and the large fragment of caspase-3 resulting from cleavage (17 kDa).


Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the cleavage site and the amino terminus of the large fragment of human caspase-3. Antibodies are purified by protein A and peptide affinity chromatography.

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).


1.  Fernandes-Alnemri, T. et al. (1994) J Biol Chem 269, 30761-4.

2.  Nicholson, D.W. et al. (1995) Nature 376, 37-43.


Entrez-Gene Id 836
Swiss-Prot Acc. P42574


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
U.S. Patent No. 5,675,063.