Custom mouse Fas ELISA Kit

Product Features

Product Name           Custom mouse Fas ELISA Kit
Assay Type                   Solid Phase Sandwich ELISA
Format                           96-well strip plate
Assay Length              4.5 hours
Sample Volume         Serum ,Plasma, cell culture media , 100ul
Detection Method    Colorimetric
Sensitivity                   This has not yet been determined
Assay Range                This has not yet been determined
Reactivity                     Mouse
Cross-Reactivity       This ELISA antibody pair only recognizes mouse Fas
Gene Names:              Fas; AI196731; APO1; APT1; CD95; TNFR6; Tnfrsf6; lpr
Gene ID                         14102
Swissprot ID               P25446

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Standards
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 µl of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4°C.
4. Wash and add 100 µl of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Wash and add 100 µl of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µl of Stop Solution to each well.
11. Read at 450 nm/570 nm.

Background

Fas, also known as APO-1, CD95, and TNFRSF6, was originally identified as a cell-surface protein which binds to monoclonal antibodies that were cytolytic for various human cell lines. Fas is highly expressed in epithelial cells, hepatocytes, activated mature lymphocytes, virus-transformed lymphocytes and other tumor cells. The ligand for Fas (FasL) has been identified and shown to be a member of the TNF family of type 2 membrane proteins. Soluble FasL can be produced by proteolysis of membrane-associated Fas. Ligation of Fas by FasL or anti-Fas antibody has been shown to induce apoptotic cell death in Fas-bearing cells. Fas plays a role in the down-regulation of the immune reaction and has been shown to be a key mediator of activation-induced death of activated T lymphocytes.

Besides the perforin/granzyme-based mechanism, the Fas system has been identified as the alternate pathway for CTL mediated cytotoxicity. FasL has also been shown to function in immunological privileged sites by killing infiltrating Fas-bearing lymphocytes and inflammatory cells.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Custom mouse Fas ELISA Kit is for Research Use Only.

• Manual
• Assay procedure
• MSDS