Glycogen Assay Kit

Glycogen Assay Kit ab65620 is an easy and accurate assay to measure glycogen levels in biological samples.

How the assay works
In the glycogen assay protocol, glucoamylase hydrolyzes the glycogen to glucose which is then specifically oxidized to produce a product that reacts with OxiRed probe to generate color (570 nm) and fluorescence (Ex 535/Em 587). The assay can detect glycogen 0.04 to 2 mg/ml.

Glycogen assay protocol summary

• - Add samples and standards to wells
• - Add hydrolysis enzyme mix and incubate for 30 min
• - Add reaction mix and incubate for 30 min
• - Analyze with microplate reader

Related Glycogen assay products
If your sample is likely to contain reducing substances, we recommend using Glycogen Assay Kit II Glycogen Assay Kit II (Colorimetric) ab169558, as reducing substances may interfere with the assay detection method used with ab65620.

Glycogen Assay Kit II (Colorimetric) ab169558 uses an alternative assay method, where glucoamylase hydrolyzes glycogen to glucose, followed by an enzymatic step which produces NADH, which is then used to reduce a tetrazolium dye, producing a colored product.

If you are running a 384 well assay, we recommend Glycogen Colorimetric Assay Kit Glycogen Colorimetric Assay Kit ab282931, which uses an identical assay method to ab65620, and is formatted for 384 well use.

How other researchers are using Glycogen Assay Kit ab65620
This glycogen assay kit has been used in publications in a variety of sample types, including:

• - Human: muscle tissue¹
• - Mouse: muscle tissue lysates², muscle and liver tissue³, liver⁴, cultured muscle myotubes⁵, astrocyte primary cell lysates⁶,
• - Rat: liver⁷, neuron-astrocyte co-cultures⁸
• - Bacteria: M. buryatense⁹, Haemophilus influenzae¹⁰

^{References: 1 - Vaughan D et al 2016, Trewin AJ et al 2015; 2 - Baligand C et al 2017, Riedl et al 2016, Wicks SE et al 2015, Todd AG et al 2015, Lundell LS et al 2019, Kim HY et al 2016, Amoasii et al 2016; 3 - Xirouchaki CE et al 2016, Pamir N et al 2015, Zachwieja NJ et al 2016; 4 - Pursell et al 2018; 5 - Park M et al 2016; 6 - Choudhury GR et al 2015; 7 - Xiang L et al 2014, Guo J et al 2018; 8 - Sobieski C et al 2018; 9 - Puri AW et al 2015; 10 - Wu S et al 2014}

Glycogen assay methods
There are 3 major methods used to assay glycogen levels:

• - Use of enzymes to produce glucose or glucose-1-phosphate from glycogen, with production of a colored reaction product (Passonneau et al 1974 PMID 4844560), such as in ab65620.
• - The anthrone method (Roe JH et al 1966, PMID 4289896) which uses alkaline digestion of glycogen, precipitation of undigested proteins with excess acid, and adds the anthrone reagent to detect glucose in a spectrophotomer. The major disadvantage of the anthrone method is the use of hazardous concentrated sulfuric acid which is heated in a >90°C water bath.
• - The phenol‐sulfuric acid method (described in Schaubroeck et al 2022, PMID 35179318), in which sulfuric acid dehydrates glycogen to 5‐hydroxymethylfurfural, which reacts with phenol to generate an orange‐colored solution, the absorbance of which can be measured spectrophotometrically. The major disadvantages of this method are the requirement to handle and resuspend pellets after centrifugation which can be fiddly, and the use of hazardous concentrated sulfuric acid and phenol.

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Other notes
This product was previously called K646 BioVision Glycogen Colorimetric/Fluorometric Assay Kit. Abcam acquired BioVision in 2021."

The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.