Apoptosis or programmed cell death is a fundamental cellular process that is essential for normal tissue development and abnormal growth such as cancer, neurodegeneration, autoimmune diseases, and angiogenesis, etc.
Apoptosis is driven by specialized proteases known as caspases.
After the initial discovery of the first mammalian caspase 1 or ICE (interleukin 1 beta converting enzyme), a growing family of caspases 1-14 have been cloned and characterized.
Caspases are synthesized as inactive zymogen or proenzyme forms (30-55kD), which upon apoptotic stimulation are proteolytically processed (self or by other proteins) in a sequential manner into their active heterotetrameric forms.
The processed form consists of large subunit (17-20kD) and a small (10-12kD) subunits, which may associate to form an active enzyme.
Functionally active caspases initiate a proteolytic cascade, capable of cleaving and activating numerous cellular targets including PARP, G4-GDI, DFF, MEKK, etc.
On a functional basis, two categories of caspases have been defined: the initiator caspases (caspases-8, -9, and –10) are activated in the earlier phases of apoptosis, whereas the executioner caspases (caspases-3, -6, and –7) are activated by initiator caspases and are responsible for dismantling cellular components.
Caspases are widely distributed in various tissues and cells.
Caspase-9 (rat/mouse 454 aa; human 416-aa, also known as apoptotic protease activating factor 3 (APAF-3)/ICE-like apoptotic protease 6 or ICE-LAP6 or MCH-6, is involved in the activation cascade of caspases responsible for apoptosis execution.
Binding of Caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3.
It is a heterodimer of a ~35kD (p35) and 10kD (p10) subunit.
Proteolytic cleavages at Asp315 by granenzyme B and at Asp330 by CPP32 generate the two active subunits.
Caspase-8 and –10 may also be involved in these processing events.
Caspase-9 is ubiquitous with highest expression in the heart, and moderate expression in liver and skeletal muscle.
Applications:Suitable for use in Western Blot and ELISA.
Other applications not tested.
Recommended Dilution:Western Blotting (1:1K-5K for neat serum and 1-10ug/ml for affinity pure antibody using ECL technique).
Caspase –9 p35 subunit is ~35 Kda
ELISA: Control peptide can be used to coat ELISA plates at 1ug/ml and detected with antibodies (1:10-50K for neat serum and 0.5-1ug/ml for affinity pure)
Optimal dilution determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
For long-term storage, store at -20°C.
Aliquots are stable for at least 12 months at -20°C.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Further dilutions can be made in assay buffer.
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Serum
Purity：Serum
Form：Supplied as a lyophilized powder from PBS, 0.05% sodium azide.
Specificity：Human control peptide is 68% conserved in rat, 65% in mouse, 64% in xenopus, and 62% in chicken caspase-9. No significant sequence homology is detected with other caspases. Since control peptide Is located at the C-terminus of p35 subunit, the antibodies will not detect the p10 subunit.
Isotype：IgG
Calc Applications Abbrev：E WB
Calc Crossreactivity：Hu
Immunogen：25-aa peptide of human Caspase-9 ~CT of p35 subunit
