ZYMUTEST™ Fibrinogen
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ZYMUTEST™ Fibrinogen (RK024A) is a two-site ELISA method for the measurement of Fibrinogen. Assay designed with affinity purified rabbit polyclonal antibodies for coating the ELISA plate and preparation of Horse Radish Peroxidase (HRP) marker. Manufactured by HYPHEN BioMed.
Intended Use
The ZYMUTEST™ Fibrinogen kit is a two site immuno-assay for measuring human Fibrinogen in plasma, or in any fluid where Fibrinogen can be present. This assay, developed with high affinity rabbit polyclonal antibodies, specific for human Fibrinogen, cross reacts with Fibrinogen from many species, and particularly with mouse Fibrinogen.
This kit is for research use only and should not be used for patient diagnosis or treatment.
Assay Principle
ZYMUTEST™ Fibrinogen is a two site ELISA, designed with rabbit polyclonal antibodies, specific for Fibrinogen.
First, the diluted tested sample is introduced into the microwells coated with rabbit polyclonal antibodies specific for Fibrinogen. When present, Fibrinogen binds onto the coated solid phase. Following a washing step, the immunoconjugate, which is a rabbit polyclonal antibody specific for Fibrinogen, coupled to horse radish peroxidase (HRP), is introduced and binds to Fibrinogen onto free epitopes. 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 color develops. When the reaction is stopped with Sulfuric Acid, a yellow color is obtained. The amount of color developed is directly proportional to the concentration of Fibrinogen in the tested sample.
Test Sample
- Human plasma collected on trisodium citrate or Na₂ EDTA anticoagulant.
- Any biological fluid where Fibrinogen must be measured.
- Citrated or Na₂ EDTA anticoagulated mouse plasma.
Reagents
- COAT: Micro ELISA plate, containing 12 strips of 8 wells, coated with a rabbit polyclonal antibody specific for Fibrinogen, then stabilized; the plate is packed in an aluminium pouch, hermetically sealed in presence of a desiccant.
- SD: 2 vials containing 60 ml of B2F-Sample Diluent, ready to use, containing a Rheumatoid factor inhibitor.
- Std: 3 vials of human Fibrinogen Standard, lyophilized. When restored with 2 ml of B2F-Sample Diluent (SD), a solution containing “C” ng/ml of human Fibrinogen is obtained (usually at about 50 ng/ml). The exact Fibrinogen concentration is indicated on the flyer provided in the kit.
- CI: 1 vial of 1 ml Fibrinogen Control I (High), lyophilized.
- CII: 1 vial of 1 ml Fibrinogen Control II (Low), lyophilized.Note: The Fibrinogen concentrations and acceptance ranges for calibrators and controls can vary from lot to lot, but are precisely indicated for each lot on the flyer provided in the kit.
- IC: 3 vials of Anti-Fibrinogen HRP immunoconjugate, a rabbit polyclonal antibody coupled to Horse-Radish-Peroxidase (HRP), lyophilized.
- CD: 1 vial of 25 ml of B2F-Conjugate Diluent, ready to use.
- WS: 1 vial of 50 ml of 20 fold concentrated Wash Solution.
- TMB: 1 vial of 25 ml peroxidase substrate: 3,3′,5,5′ – Tetramethylbenzidine containing hydrogen peroxide. Ready to use.
- SA: 1 vial of 6 ml of 0.45M Sulfuric acid (Stop solution). Ready to use.Note: Use only components from a same lot number of kits. Do not mix components from different lots of kits when running the assay.
*Use of a Rheumatoid factor Inhibitor in the sample diluent avoids the interference of Rheumatoid Factor
