Key facts
Detection method:Colorimetric/Fluorometric,
Sample types:Urine, Tissue, Cell culture supernatant, Other biological fluids,
Assay type:Quantitative,
Sensitivity:> 0.04 µg/mL,
Range:40 - 2000 ng/well,
Assay time:1h,
Assay Platform:Microplate reader
Product details:
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
ab169558
, as reducing substances may interfere with the assay detection method used with ab65620.
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
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 tissue
- Mouse: muscle tissue lysates
, muscle and liver tissue
, liver
, cultured muscle myotubes
, astrocyte primary cell lysates
- Rat: liver
, neuron-astrocyte co-cultures
- Bacteria: M. buryatense
, Haemophilus influenzae
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.
REACH authorisation
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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Glycogen is a large branched polymer of glucose serving as a form of energy storage in animals and fungi. It is sometimes called animal starch. The molecular mass of glycogen varies but it's typically around 1000000 Da. Glycogen is stored mainly in the liver and muscles. In the liver it acts as a glucose reservoir for the body while in muscles it provides energy during physical activity. In glycogen studies the use of glycogen assays glycogen measurement techniques and glycogen kits is common for quantifying its levels.
Biological function summary
Glycogen plays an important role in energy homeostasis by storing glucose that can be rapidly mobilized. Glycogen synthase and glycogen phosphorylase are critical enzymes that synthesize and degrade glycogen respectively. Glycogen synthesis and degradation are tightly regulated processes that occur in response to hormonal signals like insulin and glucagon. Glycogen often interacts with proteins that are part of complexes regulating its breakdown ensuring glucose availability when needed by the cells. Scientists often use glycogen assay kits to measure glycogen levels and the effects of its hydrolysis.
Pathways
Glycogen metabolism fits into key metabolic processes such as glycolysis and gluconeogenesis. In glycolysis the breakdown of glycogen provides glucose-1-phosphate which can further undergo conversion to glucose-6-phosphate entering the glycolytic pathway. Glycogen is also related to glycogenin a protein essential for glycogen synthesis acting as a primer for chain elongation. Within gluconeogenesis glycogen contributes glucose units for the formation of new glucose molecules maintaining blood sugar levels during fasting conditions.
Glycogen storage diseases and type 2 diabetes involve glycogen-related dysfunctions. Glycogen storage diseases are a group of metabolic disorders related to defects in glycogen synthase and glycogen phosphorylase leading to abnormal glycogen accumulation or structure. Type 2 diabetes is associated with impaired glycogen synthesis and breakdown often linked to insulin resistance. Understanding the role of glycogen in these conditions helps uncover therapeutic targets with studies utilizing glycogen tests and specific assays like those involving k648.
