Serine transhydroxymethylase: a simplified radioactive assay; purification and stabilization of enzyme activity employing Affi-Gel Blue.
An improved radioactive assay has been developed for serine transhydroxymethylase. This assay involves the direct measurement of the [14C]HCHO which is generated when [3- 14C]-serine is employed as the substrate. The new assay eliminates the need for a solvent extraction of a [14C]HCHO-dimedon adduct which is the basis of the assay devised by Taylor and Weissbach. The enzyme has been purified employing Affi-Gel Blue. The purified enzyme retains full activity when bound to this affinity chromatography matrix and can be stored in this state at 4 degrees indefinitely.
D000818:Animals; D000873:Anthracenes; D002250:Carbon Radioisotopes; D002846:Chromatography, Affinity; D002850:Chromatography, Gel / Q000379:methods; D012696:Glycine Hydroxymethyltransferase / Q000032:analysis* / Q000302:isolation & purification; D007202:Indicators and Reagents; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D011865:Radioisotope Dilution Technique; D013552:Swine; D014166:Transferases / Q000032:analysis*; D014227:Triazines
D000873:Anthracenes; D002250:Carbon Radioisotopes; D007202:Indicators and Reagents; D014227:Triazines; C012842:Cibacron Blue F 3GA; D011732:Pyridoxal Phosphate; D014166:Transferases; D012696:Glycine Hydroxymethyltransferase