Fibrinogen assay

摘要

The present invention is directed to a method of detecting intact fibrinogen, comprising the steps of: a) providing a sample containing at least some fibrinogen optionally converted at least in part to fibrin, and optionally containing thrombin; b) solubilizing the sample in a solubilizing solution that inhibits thrombin activity; c) after optional SDS-PAGE transferring/applying a portion of said sample to a protein-binding membrane; d) reacting the fibrinogen with a primary monoclonal antibody capable of binding to fibrinopeptide A moiety; and e) detecting the quantity of intact fibrinogen in the sample by quantifying the amount of the bound primary monoclonal antibody.

主权项

1. A method of detecting intact fibrinogen, comprising the steps of:
a) Providing a sample containing at least some fibrinogen optionally converted at least in part to fibrin, and optionally containing thrombin; b) Solubilizing the sample in a solubilizing solution that inhibits thrombin activity; c) Transferring a portion of said sample to a gel and subjecting said portion to electrophoresis; d) Transferring a fraction of said portion of the sample from said gel to a protein-binding membrane and immobilizing the fibrinogen on said membrane; e) Subjecting said membrane to at least one blocking step; f) Reacting the fibrinogen immobilized on said membrane with a primary monoclonal antibody capable of binding to fibrinopeptide A moiety to form a fibrinogen-antibody complex; g) Subjecting said membrane to at least one washing step to remove any unbound primary monoclonal antibody; h) Reacting said monoclonal antibody that formed the fibrinogen-antibody complex with a secondary antibody having a marker; i) Subjecting said membrane to at least one washing step to remove any unbound secondary antibody having a marker; and j) Detecting the quantity of intact fibrinogen in the sample by quantifying the amount of the marker.