Glucose 6-phosphate (G6P) is a key metabolic intermediate for glucose to enter either metabolic pathways or storage. G6P is generated when glucose is phosphorylated by hexokinase / glucokinase, or by the conversion of glucose-1-phosphate by phosphoglucomutase during glycogenolysis. G6P lies at the beginning of both glycolysis and the pentose phosphate pathways. In addition to these metabolic pathways, glucose 6-phosphate may also be converted to glycogen or starch for storage. G6P is utilized by glucose-6-phosphate dehydrogenase (G6PDH) to generate reducing equivalents in the form of NADPH. This is particularly important in red blood cells where a G6PDH deficiency leads to hemolytic anemia.

Glucose-6-Phosphate Assay Kit provides a simple, reproducible, and sensitive tool of measuring glucose-6-phosphate concentration in plasma, serum, cell lysates, and other biological liquid samples. In this assay, glucose-6-phosphate is oxidized by glucose-6-phosphate dehydrogenase, the formed NADPH is coupled to the colorimetric probe that produces a colored product which can be measured at 450 nm. The optical density measured is directly proportional to the concentration of glucose-6-phosphate present in the sample.

• Microplate reader capable of measuring absorbance at 450 nm
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Tubes to prepare sample dilutions
• 50 mL conical tubes
• MilliQ water or other type of double distilled water (ddH₂O)

1. Prepare the Reaction Mix, and Background Mix before use.
2. Add the diluted Glucose-6-Phosphate Standards or samples to the 96-well microtiter plate.
3. Initiate the reaction by adding Reaction Mix to Standards and Sample wells, and Background Mix to Sample Background wells.
4. Carefully shake the plate for a few seconds to mix.
5. Incubate the plate for 30 minutes at room temperature with mixing in the dark.
6. Measure the absorbance at 450nm using a plate reader.