active Caspase 3 antibody (ab2302)

Product Name 

active Caspase 3 antibody

Product type 

Primary antibodies

Description 

Rabbit polyclonal to active Caspase 3

Immunogen 

Synthetic peptide corresponding the cleavage site of human caspase-3.(Peptide available as ab38283.)

Reacts with 
(species key)

Hu, Ms, Rat, Ql

Specificity 

The antibody preferentially recognizes the p17 fragment of the active Caspase 3 (under the condition we tested, the antibody did not detect the precursor form and based on the purification method we used the antibody should not detect any precursor form).

Tested applications 
(see key)

ICC/IF, IHC-FoFr, IHC-Fr, IHC-P, WB

Abreviews 

Customer reviews (see key)

Read about Abreviews (Read all Abreviews)

Application notes 
(see key)

Recommended dilutions
IHC-FoFr: Use at an assay dependent dilution (PMID 18797916).
IHC-P: Suggested working dilution of 1/50 (see PMID: 16202037). In addition, found to work at 1/100 (see PMID: 15885730). For antigen unmasking, heat sections in 10 mM sodium citrate buffer (pH 6.0) for 1 minute at full power followed by 9 minutes at medium power. (Keep slides fully immersed in buffer and maintain temperature at or just below boiling.) Cool slides for 20 minutes after antigen unmasking.
IHC-Fr: Use at a concentration of 10 - 20 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 17-20 kDa.
ICC/IF: 1/200 (From PubMed:17003475).

For a positive signal the cells or tissues need to contain apoptotic cells and caspase-3 has to be activated. The recommended dilutions provide only guidelines. More antibody should be applied if very little caspase-3 has been expressed.


Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.

Positive control 
(see definition)

Rat brain,rat foetus, mouse foetus (day 9)(head, heart) (see reference Mirkes et al).
Camptothecin (2 µM) treated Jurkat cells.
If no signal is observed a time course may be required to identify maximal caspase activity.

Cellular localization 

Cytoplasmic

Research areas 

Neuroscience >> Neurology process >> Neurodegenerative disease >> Other
Cell Biology >> Apoptosis >> Intracellular >> Caspases etc >> Caspases
Neuroscience >> Neurology process >> Neurodegenerative disease >> Alzheimer's disease >> Other

Relevance 

The caspase family of cysteine proteases play a key role in apoptosis. Caspase 3 is the most extensively studied apoptotic protein among caspase family members. Caspase 3 is synthesized as inactive pro enzyme that is processed in cells undergoing apoptosis by self proteolysis and/or cleavage by other upstream proteases (e.g. Caspases 8, 9 and 10). The processed form of Caspase 3 consists of large (17kDa) and small (12kDa) subunits which associate to form an active enzyme. Caspase 3 is cleaved at Asp28 Ser29 and Asp175 Ser176. The active Caspase 3 proteolytically cleaves and activates other caspases (e.g. Caspases 6, 7 and 9), as well as relevant targets in the cells (e.g. PARP and DFF). Alternative splicing of this gene results in two transcript variants which encode the same protein. In immunohistochemical studies Caspase 3 expression has been shown to be widespread but not present in all cell types (e.g. commonly reported in epithelial cells of skin, renal proximal tubules and collecting ducts). Differences in the level of Caspase 3 have been reported in cells of short lived nature (eg germinal centre B cells) and those that are long lived (eg mantle zone B cells). Caspase 3 is the predominant caspase involved in the cleavage of amyloid beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease.

Caspases are synthesized as inactive pro-enzymes that are processed to active form in cells undergoing apoptosis. Caspase 3 has been extensively studied and implicated to play an important role in apoptosis. Active caspase 3 proteolytically cleaves and activates other caspases, as well as relevant targets in the cells (e.g., PARP). This affinity purified antibody recognizing the active forms of caspase-3 provides a new tool for identifying apoptotic cell populations in both tissue sections and cultured cells.

Database links 

Raised in 

Rabbit

Clonality 

Polyclonal

Isotype 

IgG

Purity 

Immunogen affinity purified

Storage buffer 

PBS with 1 mg/ml BSA, 1.5 mM sodium azide and 50% glycerol

Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS

Form 

Liquid

Concentration 

0.200 mg/ml

Storage instructions 

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

See below for active Caspase 3 antibody images, references, products related to ab2302 and other tools.

active Caspase 3 antibody images:

References for active Caspase 3 antibody (ab2302)

  This product has been used in: (two most recent references)

Martinez EC et al. Ascorbic acid improves embryonic cardiomyoblast cell survival and promotes vascularization in potential myocardial grafts in vivo. Tissue Eng Part A 16:1349-61 (2010). ICC/IF; Rat. PubMed: 19908964

Zehir A et al. Dicer is required for survival of differentiating neural crest cells. Dev Biol 340:459-67 (2010). PubMed: 20144605

See all 42 publication references for this product.


If you publish research using ab2302 please let us know so that we can cite the reference on this datasheet.

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