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The seriously acclaimed laboratory normal, Methods in Enzymology, is without doubt one of the so much hugely revered courses within the box of biochemistry. due to the fact 1955, each one quantity has been eagerly awaited, often consulted, and praised via researchers and reviewers alike. The sequence includes a lot fabric nonetheless proper this present day - really a necessary ebook for researchers in all fields of lifestyles sciences
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Additional info for Vitamins and Coenzymes Part G
Sample text
Lo Immediately after electrophoresis is completed, stir the gels in 500 volumes of deionized Hz0 for 15 min. Repeat the washing for two additional 15min periods. Bathe the gels in at least 2 volumes of pantetheine, dithiothreitol, cresol red, and HZ0 (1: 1: 5 : 13) and add a minimum amount of sodium hydroxide to keep the solution red in color. After lo-60 min, bands of activity in the gels will turn yellow and can be monitored at 435 nm. This procedure has also been used with l- or 3-mm slab gels.
V. Dargar, Anal. Biochem. 97, 103 (1979). E81 ACETYL PHOSPHATE AND ACETYL-COfi DETERMINATION 47 with 50 ~1 of 40 miV dithiothreitol and an additional 5-min incubation at 37”. Conversion of Acetyl-CoA to Acetyl Phosphate.
R. Cooper and J. H. Pincus, J. Agric. Food Chem. 20,490 (1972). 33 ENZYMATIC ASSAY FORPANTOTHENIC ACID [61 [61 Pantothenase-Based Assay of Pantothenic By R. ‘~* The basic reaction of this enzyme is the hydrolysis of pantothenic acid to form p-alanine and D-pantoyl lactone. Catalysis occurs through an acyl enzyme intermediate. The first acylation step is fully reversible and fast enough to enable the exchange of the P-[14C]alanine into pantothenate. p-&mine D-Pantothenate + E -zGL a pantoyl-E ti D-pantoyl lactone The rate of exchange of /3-[14C]alaninedepends on the amount of pantothenate at nonsaturating substrate concentrations, thereby making the assay possible.
