Human Caspase 8/CASP8 ELISA Kit PicoKine®

Caspase-8 is a caspase protein, encoded by the CASP8 gene. It is mapped to human chromosome 2q33-q34 and mouse chromosome 1B-proximal C. 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 composed of a prodomain, a large protease subunit, and a small protease subunit. Activation of caspases requires proteolytic processing at conserved internal aspartic residues to generate a heterodimeric enzyme consisting of the large and small subunits. This protein is involved in the programmed cell death induced by Fas and various apoptotic stimuli. The N-terminal FADD-like death effector domain of this protein suggests that it may interact with Fas-interacting protein FADD. And this protein was detected in the insoluble fraction of the affected brain region from Huntington disease patients but not in those from normal controls, which implicated the role in neurodegenerative diseases. Many alternatively spliced transcript variants encoding different isoforms have been described, although not all variants have had their full-length sequences determined.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 89 99 100 96 96 4.3 4.4% Sample 2 257 285 267 304 278 17.93 6.4% Sample 3 911 915 904 795 881 49.95 5.6% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 89 99 100 96 96 4.3 4.4% Sample 1 89 99 100 96 96 4.3 4.4% Sample 2 257 285 267 304 278 17.93 6.4% Sample 2 257 285 267 304 278 17.93 6.4% Sample 3 911 915 904 795 881 49.95 5.6% Sample 3 911 915 904 795 881 49.95 5.6%

Caspase-8; CASP-8;3.4.22.61; Apoptotic cysteine protease; Apoptotic protease Mch-5; CAP4; FADD-homologous ICE/ced-3-like protease; FADD-like ICE; FLICE; ICE-like apoptotic protease 5; MORT1-associated ced-3 homolog; MACH; Caspase-8 subunit p18; Caspase-8 subunit p10; CASP8; MCH5

Isoform 1, isoform 5 and isoform 7 are expressed in a wide variety of tissues. Highest expression in peripheral blood leukocytes, spleen, thymus and liver. Barely detectable in brain, testis and skeletal muscle.

Metabolism, Pathways and Processes, Apoptosis, Cancer, Cell Biology, Intracellular, Cell Death, Metabolism Processes, Proteolysis/Ubiquitin, Invasion/Microenvironment, Caspases, Proteolytic Enzymes, Apoptotic Markers

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 89 257 911 90 251 854 Standard deviation 6.49 16.44 46.46 6.57 20.58 46.11 CV (%) 6.4% 6.4% 5.4% 7.3% 8.2% 5.4% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 89 257 911 90 251 854 Standard deviation 6.49 16.44 46.46 6.57 20.58 46.11 CV (%) 6.4% 6.4% 5.4% 7.3% 8.2% 5.4% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 89 257 911 90 251 854 Mean (pg/ml) 89 257 911 90 251 854 Standard deviation 6.49 16.44 46.46 6.57 20.58 46.11 Standard deviation 6.49 16.44 46.46 6.57 20.58 46.11 CV (%) 6.4% 6.4% 5.4% 7.3% 8.2% 5.4% CV (%) 6.4% 6.4% 5.4% 7.3% 8.2% 5.4%

Most upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death- inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp-|-AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoform 5, isoform 6, isoform 7 and isoform 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex. .

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.019 0.08 0.131 0.219 0.437 0.808 1.531 2.62 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.019 0.08 0.131 0.219 0.437 0.808 1.531 2.62 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.019 0.08 0.131 0.219 0.437 0.808 1.531 2.62 O.D. 0.019 0.08 0.131 0.219 0.437 0.808 1.531 2.62

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.