Kit and method for measuring content of 1,3-diphosphoglycerate based on micromethod

Abstract

The invention discloses a kit and a method for measuring the content of 1,3-diphosphoglycerate based on a micromethod. A reagent of the kit comprises an extracting solution, a first reagent and a second reagent, wherein the extracting solution is prepared from Tris-HCl, EGTA, mercaptoethanol and glycerinum in a mixed mode, the first reagent is prepared from MgSO4.7H2O, NADH, ATP and glyceraldehyde-3-phosphate dehydrogenase in a mixed mode, and the second reagent is prepared from Tris-HCl. The method comprises the principle that the glyceraldehyde-3-phosphate dehydrogenase can reversely catalyze the 1,3-diphosphoglycerate and the NADH to generate glyceraldehyde-3-phosphate, inorganic phosphorus and NAD, the glyceraldehyde-3-phosphate dehydrogenase and the NADH are added into a reaction system, and by measuring the reduction amount of the NADH at 340 nm, the content of the 1,3-diphosphoglycerate can be reflected. The kit and the method for measuring the content of the 1,3-diphosphoglycerate have the characteristics that the application range is wide, operation is easy and convenient, the detection time is short, detection sensitivity is high, the cost is low, repeatability is good, and the accuracy rate is high.

Description

technical field

[0001] The invention belongs to the field of life sciences, and in particular relates to a 1,3-diphosphoglyceric acid content determination kit and a method based on a trace method. Background technique

[0002] 1,3-diphosphoglycerate is an intermediate metabolite in the glycolysis process, which is closely related to the gluconeogenesis pathway, the maintenance of blood sugar concentration in the body and the occurrence of diabetes, and plays an important role in the body's sugar, lipid and protein metabolism disorders.

[0003] At present, the existing method for detecting the content of 1,3-diphosphoglycerate is double-antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA), but this method has long detection cycle, high cost, narrow application range, poor stability and low accuracy And false positives and other shortcomings, which lead to the fact that there is no effective biochemical method to detect the content of 1,3-diphosphoglycerate i...