Invitrogen™ Human Caspase 8/FLICE ELISA Kit

Description

Includes

Aluminium pouch(es) with a Microwell Plate coated with monoclonal antibody to human Caspase-8
anti-human Caspase-8 polyclonal (rabbit) Detection Antibody
Anti-rabbit-IgG-HRP
Human Caspase-8 Standard lyophilized, 20.00ng/mL upon reconstitution
Sample Diluent
Assay Buffer Concentrate 20x (PBS with 1% Tween 20 and 10% BSA)
Wash Buffer Concentrate 20x (PBS with 1% Tween 20)
Lysis Buffer 10x
Substrate Solution (tetramethyl-benzidine)
Stop Solution (1M Phosphoric acid)
Adhesive Films

ELISA

The Human Caspase 8/FLICE ELISA quantitates Hu Caspase 8 in human serum, cell lysate or cell culture medium. The assay will exclusively recognize both natural and recombinant Hu Caspase 8. Principle of the method The Human Caspase 8 solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Caspase 8 (CASP8) is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases play a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes composed of a pro-domain, 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. Caspase 8 is involved in the programmed cell death induced by Fas and various apoptotic stimuli. The N-terminal FADD-like death effector domain of Caspase 8 suggests that it may interact with Fas-interacting protein FADD. Caspase 8 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. Caspase 8 binds to the death effector domain (DED) of FADD through an analogous DED domain present in tandem in the pro-form of the Caspase 8 protein. Activated Caspase 8 then activates other downstream caspases including Caspase 9, thereby committing the cell to undergo apoptosis. In addition, Caspase 8 also reacts with Jurkat cells and Tonsil. Overexpression of Caspase 8 induces apoptosis, which can be blocked by inhibitors specific for the ICE family. Many alternatively spliced transcript variants encoding different isoforms have been described for Caspase 8, however, not all variants have had their full-length sequences determined.

Specifications

• Accession Number: Q14790
• Assay Range: 0.16 to 10.0 ng/mL
• Assay Sensitivity: 0.10 ng/mL
• Conjugate: HRP
• Product Type: ELISA
• Sample Type: Cell Lysates, Serum, Supernatant
• Target Species: Human
• For Use With (Application): ELISA
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 841
• Gene Symbol: ALPS2B,CAP4,Casp-8,FLICE,MACH,MCH5
• Interassay CV: 8.5%
• Intraassay CV: 6.7
• Kit Contents: Pre-coated 96 well plate, Standard, Sample Diluent, Assay Buffer concentrate, Detection Antibody, anti-rabbit HRP conjugate, Lysis Buffer, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers
• Label or Dye: HRP
• Quantity: 10 x 96 Tests
• Regulatory Status: RUO
• Sample Volume: Cell Lysate 50 μL, Serum 50 μL, Supernatant 50 μL
• Storage Requirements: 2°C to 8°C
• Synonym: MACH, FADD-homologous ICE/CED-3-like protease
• Target: Caspase 8
• Test Time: 1 hrs 20 mins
• Total Assay Time: 3 hrs 15 mins

Safety and Handling

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