Product Introduction
G6P is a molecule generated by phosphorylation of the hydroxyl group on the 6th carbon of glucose under the catalysis of hexokinase. It is a common small molecule in sugar metabolism in cells and participates in biochemical pathways such as glycolysis (glycolytic pathway) and pentose phosphate pathway (PPP). In the first step of glycolysis, glucose is catalyzed by hexokinase to produce glucose-6-phosphate, and then catalyzed by phosphoglucose isomerase to form fructose-6-phosphate to continue the other steps of glycolysis; in the pentose phosphate pathway, glucose-6-phosphate is its first substrate, and this process is also the main pathway for generating NADPH. In addition to these two metabolic pathways, glucose-6-phosphate can also be converted into glycogen or starch and stored.
Product Features
1.WST-8 is an upgraded substitute for MTT, and has obvious advantages over MTT or other MTT-like products such as XTT, MTS, etc. First, the formazan generated by the reduction of MTT by some dehydrogenases is not water-soluble and requires a specific dissolving solution to dissolve; while the formazan generated by WST-8, XTT, and MTS are all water-soluble, which can save the subsequent dissolution step. Secondly, the formazan generated by WST-8 is more soluble than the formazan generated by XTT and MTS. Thirdly, WST-8 is more stable than XTT and MTS, making the experimental results more stable.
2.Compared with MTT, XTT, etc., WST-8 has a wider linear range and higher sensitivity.
3.Compared with WST-1, WST-8 has higher detection sensitivity, is more soluble, and is more stable.
4.This kit is easy to use, highly sensitive, and has a wide linear range. It can detect G6P as low as 20μM (1nmol), and shows a good linear relationship between 20 μM (1 nmol) and 500 μM (25 nmol). The detection can be performed using lysate of cells, tissues, etc., without the need to separate and purify G6P in cells, tissues, or other samples.
Principle
G6P is oxidized to generate 6-PG under the action of G6PDH. In this reaction process, NADP+ is reduced to NADPH. The generated NADPH reduces WST-8 to generate orange-yellow formazan under the action of the electron coupling reagent 1-mPMS (1-Methoxy-5-methylphenaziniumMethylSulfate), with a maximum absorption peak at around 450nm. The formazan generated in the reaction system is directly proportional to the activity of G6PDH in the sample. The principle of WST-8 method for detecting G6P is shown in the figure below:
Components
No. | Components | Size (100T) | Storage |
Reagent 1 | Enzyme Mixture | 220 μl | -20℃ |
Reagent 2 | G6P Standard (10mM) | 100 μl | -20℃ |
Reagent 3 | Color Development Solution | 220 μl | -20℃, protect from light. |
Reagent 4 | Reaction Buffer | 5.5 ml | -20℃ |
Reagent 5 | G6P Extract | 50 ml | -20℃ |
Consumable 1 | Microplate | 1 plate | RT |
Consumable 2 | Plate Sealer | 2 pieces | RT |
Storage
The unopened kit can be stored at -20°C for 12 months.
Notes
This product is intended for scientific research use only by professionals and must not be used for clinical diagnosis or treatment, in food or drugs, or stored in ordinary residences.
