Description
The method employs sandwich ELISA technique. Monoclonal antibodies are pre-coated onto microwells. Samples and standards are pipetted into microwells and Human Tissue Factor pathway Inhibitor, TFPI present in the sample are bound by the antibodies. Biotin labeled antibody is added and followed by HRP (horseradish peroxidase) conjugated secondary antibody is pipetted and incubated to form a complex. After washing microwells in order to remove any non-specific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Human Tissue Factor pathway Inhibitor, TFPI in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm