Glucose, a monosaccharide (or simple sugar), is the most important carbohydrate in biology. It serves as a vital energy source in plants, prokaryotes, and eukaryotes through mechanisms like respiration and fermentation. Glucose levels are tightly regulated in the human body. Failure to maintain blood glucose in the normal range leads to conditions of persistently high (hyperglycemia) or low (hypoglycemia) blood sugar levels. Diabetes mellitus, characterized by sustained hyperglycemia, is a prominent disease associated with impaired blood sugar regulation.

RayBio® Glucose Colorimetric Assay Kit provides a simple, reproducible, and sensitive tool for measuring glucose concentration in plasma, serum, cell lysates, urine and other biological liquid samples. This assay employs a coupled enzymatic reaction system. First, glucose is oxidized by glucose oxidase to form gluconate and hydrogen peroxide (H2O2). H2O2, in the presence of horseradish peroxidase, then reacts with a colorimetric probe to form a pink-colored product. The optical density measured at 500nm is directly proportional to the glucose concentration in the sample.

• Microplate reader capable of measuring absorbance at 340 nm at 37°C
• Precision pipettes to deliver 2 µl to 1 ml volumes
• Multi-channel pipettes to deliver 20 μl to 200 μl volumes
• Tubes to prepare sample dilutions
• Incubator at 37°C.

Positive control, sample and Sample Buffer (used as a blank) should be assayed in duplicate or triplicate. A freshly prepared positive control should be used each time the assay is performed.

1. Pre-warm the Enzyme Mix Solution at 37°C (in the dark) for at least five minutes.
2. Add 30 μL of the diluted Glucose standards or samples to the 96-well Microplate.
3. Initiate the reaction by adding 150 μL Enzyme Mix Solution to each well (pre-heat reagent at 37°C).
4. Cover with the plate cover. Carefully shake the plate for a few seconds to mix.
5. Incubate the plate for 10 minutes at 37oC in the dark.
6. Measure the absorbance at 500nm using a plate reader.