Abstract
A fluorimetric method to estimate erythrocyte glucose phosphate isomerase is described. Five μl of blood is added to 100 μl of water. Ten μl of the resulting hemolysate is incubated with a reaction mixture (200 μl) containing Tris-HCl buffer pH 8.0, 20 μmoles, MgCl2 2 μmoles, nicotinamide adenine dinucleotide phosphate 0.08 μmoles, glucose-6-phosphate dehydrogenase 0.2 EU and fructose-6-phosphate 0.12 μmoles. After ten minutes of 25°C 20 μl of the mixture is added to 2 ml of 0.01 M phosphate buffer pH 7.4 and the nicotinamide adenine dinucleotide phosphate fluorescence determined. Two ml of water was added to the remaining reaction mixture and the hemoglobin concentration determined at 410 nm. The technique is primarily designed for use with small amounts of blood, of widely varying activity, from various animal species.
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Selected References
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