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Glycogen Assay Kit ab65620 is a no-wash glycogen assay with two 30-minute incubation steps. In the assay, glucoamylase acts on glycogen, and the resulting glucose is oxidized, with readout on colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) plate reader.

Convenient glycogen assay kit designed for success:

  • - Complete kit format with full assay protocol and standard curve for quantitation
  • - Published extensively with tissue lysates (liver, muscle, other) and cultured cell lysates
  • - Flexible format with individual kit components also available for purchase (with a minimum order of 20 units; contact us to discuss your needs)

Is this your first time purchasing this assay kit? Get 20% off one kit by  entering code 20OFF-T5A8Q at checkout. T&C’s apply – click here. Offer only valid when purchasing directly from Abcam in Europe, North America, Australia, and Singapore.

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Key facts

Detection method

Colorimetric/Fluorometric

Sample types

Urine, Tissue, Cell culture supernatant, Other biological fluids

Assay type

Quantitative

Range

40 - 2000 ng/well

Assay time

1h

Sensitivity

> 0.04 µg/mL

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What's included?

2000 Test
Components
Assay Buffer II
20 x 25 mL
Assay Buffer VIII
20 x 25 mL
Development Enzyme Mix II
20 x 1 Vial
Glycogen Standard
20 x 100 µL
Hydrolysis Enzyme Mix I
20 x 1 Vial
OxiRed Probe
20 x 200 µL

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Glycogen Assay Kit ab65620 is a no-wash glycogen assay with two 30-minute incubation steps. In the assay, glucoamylase acts on glycogen, and the resulting glucose is oxidized, with readout on colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) plate reader.

Convenient glycogen assay kit designed for success:

  • - Complete kit format with full assay protocol and standard curve for quantitation
  • - Published extensively with tissue lysates (liver, muscle, other) and cultured cell lysates
  • - Flexible format with individual kit components also available for purchase (with a minimum order of 20 units; contact us to discuss your needs)

Is this your first time purchasing this assay kit? Get 20% off one kit by  entering code 20OFF-T5A8Q at checkout. T&C’s apply – click here. Offer only valid when purchasing directly from Abcam in Europe, North America, Australia, and Singapore.

Key facts

Detection method

Colorimetric/Fluorometric

Sample types

Urine, Tissue, Cell culture supernatant, Other biological fluids

Assay type

Quantitative

Range

40 - 2000 ng/well

Assay time

1h

Assay Platform

Microplate reader

Sensitivity

> 0.04 µg/mL

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

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

Which glycogen assay kit should I choose?
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 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-related diseases and pathways, and supporting research tools
Glycogen is a multibranched polysaccharide of glucose, that is a core energy reservoir in liver and muscles.

Glycogen is involved in several metabolic diseases:

- Glycogen Storage Diseases are inherited disorders caused by enzyme deficiencies affecting glycogen metabolism; von Gierke, Cori/Forbes, and Pompe disease are three of the most common.
Use glucose-6-phosphatase antibodies and glucose-6-phosphate assay kits to study the enzyme deficient in von Gierke Disease. Use alpha glucosidase assay kits to study the enzyme deficient in Pompe Disease.

- Type II Diabetes, metabolic syndrome, insulin-resistance, and non-alcoholic fatty liver disease are all connected to obesity, and are related to glycogen storage.
Use insulin ELISA kits and glucagon ELISA kits to study key hormones involved in these diseases. Review tools to study fatty liver, liver fibrosis, and liver toxicity.

The major pathways for glycogen synthesis and glycogen breakdown are glycogenesis and glycogenolysis:
Study key enzymes involved in these pathways with glycogen synthase assay kits and antibodies, glucose-6-phosphate assay kits, and glycogen phosphorylase assay kits and antibodies.

Review our metabolism assays guide to learn about the assay kits we manufacture for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress.

Alternative glycogen assay methods
There are 3 major methods uses 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 >90oC 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.

Regulatory notes

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.

Supplementary info

Biological 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.

Mechanical summary

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.

Pathway

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.

Disease

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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4 product images

  • Functional Studies - Glycogen Assay Kit (ab65620), expandable thumbnail
    Image from Roy Choudhury G et al., PLoS One 10(4), Fig 6c. Doi: 10.1371/journal.pone.0123096. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Functional Studies - Glycogen Assay Kit (ab65620)

    Total glycogen levels in C576bL6 mice astrocytes were determined by using Glycogen assay kit (ab65620). At 24 hours following OGD-reoxygenation, astrocytes had less glycogen levels compared to normoxia control. Astrocytes treated with Methylene blue (MB) showed a higher glycogen content compared to non-MB treated, OGD-reoxygenation astrocytes. * p < 0.05; ## p < 0.001 Vs. OGD-reoxygenation control / 0 μM MB.

  • Functional Studies - Glycogen Assay Kit (ab65620), expandable thumbnail

    Functional Studies - Glycogen Assay Kit (ab65620)

    Functional Studies - Glycogen Assay Kit.

    Example of fluorometric standard curve using Glycogen Assay Kit (ab65620).

  • Functional Studies - Glycogen Assay Kit (ab65620), expandable thumbnail

    Functional Studies - Glycogen Assay Kit (ab65620)

    Measurement of glycogen in various mouse tissues using Glycogen Assay Kit (ab65620).

  • Functional Studies - Glycogen Assay Kit (ab65620), expandable thumbnail

    Functional Studies - Glycogen Assay Kit (ab65620)

    Glycogen concentration measured in MBA-MB-231 cells (human breast adenocarcinoma cell line). 106 cells were prepared following protocol instructions, and several dilutions were measured using fluorometric detection.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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