Caspase-3 is a caspase protein that is encoded by the CASP3 gene. This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein cleaves and activates caspases 6, 7 and 9, and the protein itself is processed by caspases 8, 9 and 10. It is the predominant caspase involved in the cleavage of amyloid-beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease. Alternative splicing of this gene results in two transcript variants that encode the same protein.
Sample Type
Cell culture supernates, serum.
Intended Use
Sandwich High Sensitivity BioAssay™ ELISA kit for Quantitative Detection of Human Caspase 3.
Range
31.2pg/ml-2000pg/ml
Specificity
Recognizes natural and recombinant Human Caspase 3. There is no detectable cross-reactivity with other relevant proteins.
Test Principle
This Human Caspase 3 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A specific for Caspase 3 has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli; Immunogen sequence: S29-D175&A183-H277) and test samples are added to the wells, a biotinylated detection specific for Caspase 3 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Human Caspase 3 amount of sample captured in plate.
Kit Components
96-well plate precoated with anti-Human CASP3 antibody, 1x plate Lyophilized recombinant Human CASP3 standard, 2x10ng/tube Biotinylated anti-Human CASP3 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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