By David Glick
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Extra resources for Methods of Biochemical Analysis, Volume 5
90 ml. of distilled water is added and then NaOH, dropwise, until the solution just turns blue. The total volume is made up to 100 ml. 7 (this should be checked, if possible, with a pH meter). For use, two volumes of this stock solution are diluted with one volume of distilled water. Procedure. (1) 20 pl. of blood is collected in a hemoglobin pipette and washed out into 1 ml. of dilute BTB solution contained in one tube. 5 ml. of ACh solution is added and the time of addition recorded. (3)The exact time at which the color of the reaction mixture becomes deep orange is noted.
The results obtained by these groups of workers (Table VI) are in most cases comparable. Modifications of Michel’s method have been proposed; the results thus obtained are not directly comparable with those obtained using the original method. I n the first place certain modifications of the buffer solutions and enzyme dilutions have been employed (Table VII). , have been published (58, 106). The method described by MacDonald el nl. (106) for the assay of erythrocyte activity in whole blood is based on the differentactivity-substrate con- 28 KLAS-BERTIL AUGUSTINSSON centration relationships of the plasma and erythrocyte enzymes.
For use, two volumes of this stock solution are diluted with one volume of distilled water. Procedure. (1) 20 pl. of blood is collected in a hemoglobin pipette and washed out into 1 ml. of dilute BTB solution contained in one tube. 5 ml. of ACh solution is added and the time of addition recorded. (3)The exact time at which the color of the reaction mixture becomes deep orange is noted. The time interval is a measure of the enzyme activity. B. 15 ml. 01 N acetic acid. 5 ml. 7). Estimation. ) against time (minutes)) is used to obtain information as to which of the thrce “zones” the observed enzyme level (in minutes) is related.
Methods of Biochemical Analysis, Volume 5 by David Glick