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Determination of 1,3-diphosphoglycerate and other intermediates in the human red blood cells by the firefly luciferase method

✍ Scribed by Günther Momsen

Publisher
Elsevier Science
Year
1977
Tongue
English
Weight
574 KB
Volume
82
Category
Article
ISSN
0003-2697

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✦ Synopsis

A highly sensitive and rapid method for the determination of 1,3-diphosphoglycerate (1,3-DPG) has been developed. The method is designed to measure 1.3-DPG concentrations as small as 10 nmol/liter of cells. The very high sensitivity of the determination of ATP by the firefly luciferase method [Rasmussen and Nielsen (1968) Acta Chetn. Scud. 22, 1745-1756;Rhee, Greifner, and Chock (1975) Anal. Biochem. 66, 259-2641 was utilized. ATP from the cells was removed with active charcoal, and in a neutralized filtrate 1.3-DPG was quantitatively converted to ATP and 3-phosphoglycerate (3-PG) by adding ADP and phosphoglycerate kinase (PGK). 493

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