|
SampleTypes |
|
plasma, serum, milk, urine, saliva, and cell culture supernatants
|
|
Protocol |
|
Human Factor X (FX) ELISA kit is designed for detection of human factor X in plasma, serum, milk, urine, saliva, and cell culture supernatants. This assay employs a quantitative sandwich enzyme immunoassay technique that measures FX in less than 4 hours. A monoclonal antibody specific for FX has been pre-coated onto a 96-well microplate with removable strips. FX in standards and samples is sandwiched by the immobilized antibody and the peroxidase conjugated polyclonal antibody specific for FX. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.
|
|
Storage |
|
Store components of the kit at 2-80C or -200C upon arrival up to the expiration date.
|
|
References |
|
(1) Ruf, W. and Edgington, T.S. (1994) FASEB J. 8:385
(2) Neuenschwander, P.F. et al. (1993) Thrombosis and Haemostasis 70:970
(3) Messier, T.L. et al. (1991) Gene 99:291
(4) Di Scipio, R.G. et al. (1977) Biochemistry 16:5253
|
|
Contents/Specifications |
|
Reagents;
• FX Microplate: A 96 well polystyrene microplate (12 strips of 8 wells) coated with a monoclonal antibody against human FX.
• Sealing Tapes: Each kit contains 3 pre-cut, pressure-sensitive sealing tapes that can be cut to fit the format of the individual assay.
• FX Standard: Plasma human FX in a buffered protein base (400 ng, lyophilized).
• Biotinylated FX Antibody (50x): A 50-fold biotinylated polyclonal antibody against H. FX (140 μl).
• Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (80 μl).
• MIX Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml).
• Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml, 2 bottles).
• Chromogen Substrate: A ready-to-use stabilized peroxidase chromogen substrate tetramethylbenzidine (8 ml).
• Stop Solution: A 0.5 N hydrochloric acid to stop the chromogen substrate reaction (12 ml).
|
|
Notes |
|
Factor X (FX) is a plasma serine protease zymogen involved in the blood coagulation cascade. FX is purified from plasma as a two-chain protein consisting of a 45 kDa heavy chain and a 17 kDa light chain. The FX heavy chain is cleaved during coagulation by several different proteases including the intrinsic Xase complex, the FX-activating enzyme from Russell’s viper venom (RVV) and trypsin, and also by extrinsic (tissue factor/factor VIIa) pathway to give an active enzyme FXa. FXa as the activator of prothrombin occupies a central position linking the two blood coagulation pathways (1 - 4).
|