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Additional info for Methods of Biochemical Analysis, Volume 21
22 DAVID ZAKlM AND DONALD A. VESSEY contain “toxins” which do not have “lecithinase” activity (32). 6, in a sealed ampoule. Approximately 50% of the phospholipase C is also lost during heating. The effects of pure preparations of phospholipase C on the properties of microsomal membranes have not been studied. Two isoenzymes of phospholipase C can be purified from extracts of Clostridium welchii (33,34); sphingomyelin is the preferred substrate for one isoenzyme, and lecithin for the other (34).
A solution to the problem of reoxidation of reduced nucleotides as a competing reaction seems to be to study reactions requiring NADH or NADPH anaerobically, since in the microsomes 0 2 is the only terminal electron acceptor. Care should be exercised in the use of known inhibitors of interfering enzymes, and the effects of the inhibitor on the activity of the enzyme of primary interest should be susceptible to investigation in an independent manner. For example, EDTA and sugar-nucleotides, other than UDPglucuronic acid, are effective inhibitors of the microsomal nucleotide pyrophosphatase (64) ; but EDTA inhibits the activity of UDP glucuronyltransferase, and UDP-N-acetylglucosameine activates this enzyme in guinea pig livers.
WHITEHEAD I. INTRODUCTION Selenium occurs naturally in biological materials at levels that vary from a few parts per billion to a few per cent. Methods for its determination in these materials must, therefore, cover a very wide range of concentrations. As a result there have been developed a number of procedures for selenium analysis, many of which are discussed briefly in a review by Watkinson (1). The methods generally used for analyzing biological materials for selenium are based on the destruction of the organic constituents with the concomitant oxidation of the element to Se4+or Se6+and its subsequent determination by one of a variety of procedures, either with or without its separation from other elements.
Methods of Biochemical Analysis, Volume 21