Purification and Characterization of Hypoxanthine-Guanine Phosphoribosyl Transferase from Cultured HTC Cells
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Hypoxanthine-guanine phosphoribosyl transferase (HGPRTase.) from |35S|-methionyl-labelled rat hepatoma (HTC) cells has been purified more than 600-fold to near homogeneity by making use of conventional and affinity chromatographic techniques (Table 1). The conventional purification procedure utilizes a pH 5.0 precipitation, an ammonium sulfate precipitation, DEAE-cellulose chromatography, and heating at 80–85°according to the method of Olsen and Milman, 1974. Using this procedure, a partially pure enzyme preparation is obtained. Affinity chromatography according to Hughes et al., 1975, can be used either before or after the DEAE step to yield pure enzyme.
KeywordsCrude Extract Pure Enzyme Preparation Enlarge Portion Potassium Phosphate Buffer 60mM Major Isoenzyme
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