Rabbit Anti-Caspase 8 (FLICE, CASP-8, FLICE, ICE-like apoptotic protease 5, MACH, MCH5) (Not for Export EU)
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-8, also known as (MACH, FADD-like ICE, FLICE, Mch-5, CAP4, MACH-alpha 1) human proenzyme is 479 aa (mouse 480 aa, rat 482 aa). It is the most upstream protease of the activation of caspases responsible for CD95 and TNFR-1 induced cell death. It cleaves and activates caspases-3, -4, -6, -7, -9, and –10. It is a heterodimer of a 19kD (p18) and 10kD (p10) subunits. Caspase-8 is alternatively spliced into at least 8 isoforms (1-4 alpha, 1-4 beta). Alpha 1 and beta-1 are expressed in a many tissues. Highest expression is observed in peripheral blood, leukocytes, spleen, thymus and liver. Lower expression is seen in brain and testis.
Applications
Suitable for use in Western Blot and ELISA. Other applications not tested.
Applications
Western Blotting (1:1K-5K using ECL technique). Caspase pro-enzyme is ~55kD that is processed to generate heterodimer of p18 and p10/12. 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).
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.
Immunogen
15-aa peptide within the p18 domain of human Caspase-8 p18 domain
Form
Supplied as a lyophilized powder from PBS, 0.05% sodium azide.
Specificity
Human control peptide is 69% conserved in mouse caspase-8. No significant sequence homology is detected with other caspases.