Glycogen Assay Kit (Colorimetric)

Glycogen is a polysaccharide found in animals as well as simpler organisms such as fungi. Glycogen is made up of glucose monomers and is the primary method of storing glucose in animals. It is stored primarily in the liver and muscle, forming an energy reserve that can be quickly mobilized to meet sudden needs for glucose. The most common glycogen metabolism disorder is found in diabetes, where abnormal insulin levels cause liver glycogen to be abnormally accumulated or depleted. Genetic glycogen storage diseases are associated with various inborn errors of metabolism caused by deficiencies of enzymes necessary for glycogen synthesis or breakdown.

Glycogen Assay Kit provides a simple, reproducible, and sensitive tool of measuring glycogen concentration in cell lysates, urine, saliva, and other biological liquid samples. This assay employs a coupled enzymatic reaction system that combines multiple steps. Initially, glycogen is hydrolyzed by amyloglucosidase to form β-D-glucose, which is then specifically oxidized by glucose oxidase to form hydrogen peroxide (H2O2). Peroxidase then catalyzes the reaction of hydrogen peroxide with the Colorimetric Probe to form a pink colored product. The optical density measured at 510nm is directly proportional to the concentration of glycogen present in the sample.

• Microplate reader capable of measuring absorbance at 510 nm
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Tubes to prepare sample dilutions
• Incubator at 37°C
• 50 mL conical tubes

1. Add 20 µL of the diluted Glycogen Standards or samples to the 96-well microtiter plate.
2. For Standard Wells and Sample Wells: add 10 µL of the Amyloglucosidase Solution (See “Reagent Preparation”) to the corresponding wells.
   For Sample Background Wells: add 10 µL of hydrolysis buffer to corresponding wells.
3. Add 150 µL of Glucose Reagent to each well. Incubate the plate at 37°C for 30 minutes with gently mixing in the dark.
4. Measure the absorbance at 510 nm using a plate reader.