The Glycerol 3-Phosphate Dehydrogenase Assay Kit is a 96-well microtiter plate assay designed to measure the activity of GPDH. This kit can be used to either screen differentiation repressors or to study mechanisms regulating differentiation of primary or established cell-line progenitor cells into adipocytes. The procedure involves extracting cells by addition of the Enzyme Extraction Buffer, followed by serial dilution of the extract. The serially diluted extract is added to a 96-well plate that contains substrate solution, and GPDH activity is calculated by measuring the change (decrease) in absorbance at OD340 using a microtiter UV plate reader.
The enzyme glycerol 3-phosphate dehydrogenase catalyzes a reversible reaction between dihydroxyacetone phosphate (DHAP) and glycerol 3-phosphate, with NAD as a coenzyme. GPDH activity increases significantly during differentiation of progenitor cells into adipocytes and, thus, the activity of this enzyme has been used as an index for monitoring fat synthesis.
