CASPASE-9 Assay Kit | CASP9 assay kit
FAM FLICA CASPASE-9 KIT
10X Wash Buffer, 15 mL
Fixative, 6 mL
Propidium Iodide, 1 mL
Hoechst Stain, 1 mL
Protect the FLICA reagent from light at all times.
Once reconstituted, the 150X FLICA stock should be stored at -20 degree C protected from light.
Shelf Life: The 150X FLICA reconstituted stock is stable for up to 6 months and may be thawed twice during that time.
Testing Data #1
(Both cells are apoptotic (green) and dying (blue nuclei). As the left cell is much brighter green than the right cell, the left cell had more active caspases)
Test Principle: Caspase FLICA kits measure apoptosis by detecting active caspases in whole, living cells. These kits do not work by using antibodies or as an ELISA. Instead, their methodology is based on a unique cell-permeable and non-cytotoxic reagent called the Fluorochrome Inhibitor of Caspases (FLICA). The FLICA reagent contains a caspase inhibitor sequence linked to a green (Carboxyfluorescein, FAM) fluorescent probe.
The Caspase FLICA Kits are suitable for cells in suspension, adherent cells, thin tissue sections (but not fixed or paraffin-embedded cells) from many species including mammalian, insect and yeast. Different cell types, e.g. HeLa, primary neurons, macrophages and lymphocytes have also been successfully studied with these kits.
This kit can be used with a flow cytometer, fluorescence microscope or a fluorescence plate reader using black microtitre plates.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes 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 can undergo autoproteolytic processing and activation by the apoptosome, a protein complex of cytochrome c and the apoptotic peptidase activating factor 1; this step is thought to be one of the earliest in the caspase activation cascade. This protein is thought to play a central role in apoptosis and to be a tumor suppressor. Alternative splicing results in multiple transcript variants. [provided by RefSeq, May 2013]
Uniprot Description
CASP9: 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 2 subunits, large and small, that dimerize to form the active enzyme. This protein is processed by caspase APAF1; this step is thought to be one of the earliest in the caspase activation cascade. Alternative splicing results in two isoforms.
Protein type: Protease; EC 3.4.22.62; Apoptosis
Chromosomal Location of Human Ortholog: 1p36.21
Cellular Component: mitochondrion; nucleus; cytosol; apoptosome
Molecular Function: peptidase activity; protein binding; cysteine-type endopeptidase activity; enzyme activator activity; protein kinase binding; SH3 domain binding
Biological Process: epidermal growth factor receptor signaling pathway; phosphoinositide-mediated signaling; fibroblast growth factor receptor signaling pathway; nerve growth factor receptor signaling pathway; positive regulation of apoptosis; apoptosis; DNA damage response, signal transduction; response to lipopolysaccharide; proteolysis; response to estradiol stimulus; response to antibiotic; DNA damage response, signal transduction resulting in induction of apoptosis; positive regulation of neuron apoptosis; response to cobalt ion; innate immune response; platelet formation; caspase activation via cytochrome c; response to DNA damage stimulus; aging
