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ZYMUTEST™ FPA (RK016B) is a Competitive Enzyme-Linked Immunosorbent Assay (CELIA) for the in vitro quantitative determination of human FPA, using on bentonite adsorbed human plasma, or in any fluid where FPA can be present. Fibrinogen, when present in the specimen, must be removed (i.e., bentonite adsorption of human plasma), as it cross-reacts with antibodies to FPA.. Manufactured by HYPHEN BioMed.
For improvement purposes HYPHEN BioMed decided to no longer offer the Elisa assay for the measurement of FPA (ZYMUTEST™ FPA – RK016A) and quality controls (FPA Control – SC015K) separately. A new “all-in-one” version of ZYMUTEST™ FPA was released on October 1st, 2020 under the product reference RK016B.
FPA is measured on bentonite adsorbed human plasma, which is then fibrinogen free. In a first step, FPA calibrator or tested sample is preincubated with a constant and limited amount of affinity purified rabbit antibodies specific for human FPA. In a second step, the unreacted anti-FPA antibodies are then measured using a micro ELISA plate coated with synthetic FPA and stabilised. Free antibodies bind to immobilised FPA. Following a washing step, the immunoconjugate, which is a goat polyclonal antibody specific for rabbit IgGs and coupled to Horse-Radish-Peroxidase (HRP), is introduced into microwells and binds to immobilised anti-FPA.
Following a new washing step, the peroxidase substrate, 3,3′,5,5′ –Tetramethylbenzidine (TMB), in presence of hydrogen peroxide (H2O2), is introduced and a blue colour develops. The colour turns yellow when the reaction is stopped with Sulfuric Acid. There is an indirect relationship between the colour developed and the concentration of FPA in the tested sample.