Download Carbohydrate Metabolism - Part D by Willis A. Wood (Eds.) PDF

By Willis A. Wood (Eds.)

The seriously acclaimed laboratory regular, Methods in Enzymology, is likely one of the such a lot hugely revered guides within the box of biochemistry. in view that 1955, each one quantity has been eagerly awaited, often consulted, and praised by way of researchers and reviewers alike. The sequence comprises a lot fabric nonetheless appropriate this day - really a necessary booklet for researchers in all fields of existence sciences

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The final a b s o r b a n c e was corrected for the a b s o r b a n c e contributed by the D - L D H in the N A D ÷, g l y c i n e hydrazine solution. 177 + , D-LDH added; - , w a t e r added in place of D-LDH. b + , P l a s m a filtrate in buffer; - , w a t e r a dde d in place of p l a s m a filtrate. c At 340 mm against a c u v e t t e containing only water. Using Eq. 25. 027 mM. 023 mM). 21 These values are in the range of 1-2% of the concentration of L-lactate. Discussion The assay described here was directed toward D-lactate concentrations that would be below those expected for the normal glycolytic product, L-lactate.

287. [4] ENZYMIC MICRODETERMINATION FOR SUBSTRATES 23 version of the method can be used. Substrate oxidation can be assayed with other methods using phenazine methosulfate or 2,6-dichlorophenolindophenol as the electron acceptor. Although such assay methods show a good agreement with each other, the ferricyanide method is mainly employed in this chapter owing to its simplicity for routine use. Further, with 2,6-dichlorophenolindophenol, the molecular extinction coefficient is variable with pH, and this makes the assay less reliable for quantitative determination.

Schwartz, G. Kessler, and O. Bodansky, J. Biol. Chem. 236, 1207 (1946). 6 j. F. Speck, J. W. Moulder, and E. A. Evans, J. Biol. Chem. 164, 119 (1946). 7 C. S. Apstein, E. Puchner, and N. Brachfeld, Anal. Biochem. 38, 20 (1970). 8 j. C. Rosenburg and B. F. Rush, Clin. Chem. 12, 299 (1969). 9 Boehringer Mannheim, Biochem. Bull. 120, 121 (1973). [5] L(-k)- AND D( -- )-LACTATE DETERMI NATION 31 TABLE I LACTIC AOD ASSAY REAGENTS Procedure Reagent No. 15% albumin (bovine fraction) L. 50 ml in 20 ml of reagent 3, providing 800 units of activity) 500 mg of NAD ÷ dry powder d (ethanol free) added to reagent 4 immediately prior to assay " Brij from Fisher Scientific Co.

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