Description
Introduction: Fibrinogen is a soluble glycoprotein that circulates in the blood and is converted to insoluble fibrin by thrombin in the final step of the coagulation cascade 1. Hepatic expression of fibrinogen increases two to four hundred fold during the acute phase response to infection or inflammation
2. Elevated fibrinogen levels are correlated with cardiovascular disease3 and atherosclerosis4.
Principle of assay: This is an ELISA (Enzyme-Linked Immunosorbent Assay) for the quantitative analysis of total fibrinogen levels in rat plasma and serum. This test kit operates on the basis of sandwich ELISA where the fibrinogen binds to a capture antibody on the plate and is quantified with the use of a biotin labeled primary antibody and an avidin-HRP conjugate.
Rat fibrinogen will bind to the affinity purified capture antibody coated on the microtiter plate. After appropriate washing steps, biotin labeled polyclonal anti-rat fibrinogen primary antibody binds to the captured protein. Excess antibody is washed away and bound polyclonal antibody is reacted with avidin conjugated to horseradish peroxidase. TMB substrate is used for color development at 450nm. A standard calibration curve is prepared along with the samples to be measured using dilutions of rat fibrinogen. Color development is proportional to the concentration of fibrinogen in the samples