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

By Willis A. Wood (Eds.)

The significantly acclaimed laboratory common, Methods in Enzymology, is among the such a lot hugely revered courses within the box of biochemistry. considering 1955, each one quantity has been eagerly awaited, usually consulted, and praised via researchers and reviewers alike. The sequence comprises a lot fabric nonetheless suitable this present day - actually a vital book for researchers in all fields of lifestyles sciences

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Gasser, Abstr. , lOth No. 604 (1975). 4 R. A. U s m a n o v and G. A. Kochetov, Biochem. Int. 3, 33 (1981). VOL. 89 Copyright© 1982by AcademicPress. Inc. All rightsof reproductionin any formreserved. ISBN 0-12-181989-2 20 ANALYTICALMETHODS [4] reaction is complete at 20°. The time is usually within 5-15 min, depending on the quantity of fructose 6-phosphate in the sample. 1 scale is used. Calculation. The molar extinction coefficient for ferricyanide at 420 nm is 1000 M -1 c m - k The quantity of fructose 6-phosphate (/zmol) is calculated from the formula M = A420/2 " V where V is the volume of the reaction mixture.

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.

L a c t a t e conc. (mg/100 ml) = change in a b s o r b a n c e - a b s o r b a n c e intercept x dilution factor slope (2) The actual values determined to express the p l a s m a concentration o f D-lactate are given in Eq. (3). L a c t a t e conc. 595 x 4 (3) Plasma D-Lactate Assay. T h e p r o c e d u r e was the s a m e as for the standard c u r v e e x c e p t that heparinized blood was centrifuged at 3200 r p m (1400 g) for 10 min in a clinical centrifuge and 2 ml o f the p l a s m a was added to the Amicon cone.

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