MEASURING METHOD FOR ACTIVITY OF CELL DIFFERENTIATION INDUCER

摘要

PURPOSE:To enable processing of multiple specimens by reducing NBT by the active oxygen produced with the differentiation of undifferentiated cells eluting the blue formazan compd. formed in the cells to the outside of the cells and measuring the blue absorbancy of the compd. thereof. CONSTITUTION:The undifferentiated cells for measurement are pretreated by actinomycin D and are cultured after the addition of a differentiation inducer thereto. A proper amt. of the NBT (nitroblue tetrazolium) reagent is added to the culture liquid and a reduction reaction is caused after stirring. A hydrochloric acid is added to the liquid to stop the reaction and thereafter the deposited cells of the blue formazan compd. are settled in a test vessel. A proper amt. of the eluting solvent is added to the cells deposited with the dyes to elute and homogenize the dyes. The light absorbancy of the dye elute is then measured. A multiscan is preferably used for the measuring instrument and the multispecimens are automatically measured by using a computer. The calculation of the activity is executed by using a standard specimen of a known activity unit and calculating the activity of the differentiation inducer of the sample to be detected by the relative degree of the light absorption.