Effect of mercaptoethanol in the radioactive thin layer chromatography assay of NAD+-15-hydroxyprostaglandin in dehydrogenase.
The use of mercaptoethanol in the assay of rat kidney 15-hydroxyprostaglandin dehydrogenase (PGDH) was found to have minimal effect on activity assayed with the spectrophotometric and substrate loss assays. However, mercaptoethanol appeared to inhibit PGDH when assayed by thin-layer chromatography, based upon conversion of 3H-PGE1 to 15-keto-3H-PGE1. Mercaptoethanol reacted with 15-keto-PGE1 to alter its chromatographic mobility and to suppress the U.V. absorption spectrum of 15-keto-PGE1. The implication of the use of ME in radiometric assays is discussed.
D000818:Animals; D002855:Chromatography, Thin Layer; D004229:Dithiothreitol / Q000494:pharmacology; D006911:Hydroxyprostaglandin Dehydrogenases / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology; D008623:Mercaptoethanol / Q000494:pharmacology*; D009249:NADP / Q000378:metabolism; D011458:Prostaglandins E / Q000378:metabolism; D011460:Prostaglandins F / Q000378:metabolism; D051381:Rats
D011458:Prostaglandins E; D011460:Prostaglandins F; D009249:NADP; D008623:Mercaptoethanol; D006911:Hydroxyprostaglandin Dehydrogenases; D004229:Dithiothreitol