An eight point standard curve is generated using the MLA 1400C. NPP Plasma (made and characterized in house) dilution is started at a 1:5 and subsequently serial diluted. To this dilution, an equal volume of FVII deficient plasma is added (1:1). The dilution is incubated at 37ºC for 106 seconds. Clotting is initiated by addition of 2 parts PT reagent [Hemoliance RecombiPlasTin; Lexington, MA]. A new standard curve was generated for each testing day.
The purified samples are prepared by making a dilutions into deficient plasma. These samples are then serially diluted in Assay Buffer by the MLA 1400C from a 5-fold dilution for a total of three dilutions. The MLA 1400C compares the clot times to the standard curve and expresses the result as a percentage on the printout.
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