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Glycogen Assay Kit (ab65620)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (45)References (46)

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Functional studies - ab65620
  • Functional Studies - Glycogen Assay Kit
  • Functional Studies - Glycogen Assay Kit (ab65620)
  • Functional Studies  - Glycogen Assay Kit (ab65620)

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric/Fluorometric
  • Platform: Microplate reader
  • Assay time: 1 hr
  • Sample type: Cell culture supernatant, Other biological fluids, Tissue, Urine
  • Sensitivity: 0.4 µg/ml

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Overview

  • Product name

    Glycogen Assay Kit
    See all Glycogen kits
  • Detection method

    Colorimetric/Fluorometric
  • Sample type

    Cell culture supernatant, Urine, Other biological fluids, Tissue
  • Assay type

    Quantitative
  • Sensitivity

    > 0.4 µg/ml
  • Range

    0.4 µg/ml - 2000 µg/ml
  • Assay time

    1h 00m
  • Product overview

    Glycogen Assay Kit ab65620 is an easy and accurate assay to measure glycogen levels in biological samples. 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 0.0004 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


    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.

  • Notes

    Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

     

    How other researchers have used Glycogen Assay Kit ab65620

    The 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

    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.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Storage buffer

    Preservative: None
    Constituents: 0.1% Triton-X-100, DMSO
  • Components Identifier 100 tests
    Development Buffer WM 1 x 25ml
    Development Enzyme Mix (lyophilized) Green 1 vial
    Glycogen Standard (2.0 mg/ml) Yellow 1 x 100µl
    Hydrolysis Buffer NM 1 x 25ml
    Hydrolysis Enzyme Mix (lyophilized) Blue 1 vial
    OxiRed Probe Red 1 x 200µl
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Sugar Assays
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Lipid Metabolism Kits
    • Insulin signaling
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Lipid Metabolism Kits
    • Obesity
  • Relevance

    Glycogen is the primary short term energy storage molecule in animals. It is synthesized primarily in the liver and muscle. Glycogen is a highly branched polymer of glucose molecules, connected with an alpha-1,4 linkage, branching via an alpha-1,6 linkage. Abnormal ability to utilize glycogen is found in diabetes and in several genetic glycogen storage diseases.

Associated products

    Images

    • Functional studies - ab65620
      Functional studies - ab65620Image 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/

      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
      Functional Studies - Glycogen Assay Kit

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

    • Functional Studies - Glycogen Assay Kit (ab65620)
      Functional Studies - Glycogen Assay Kit (ab65620)

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

    • Functional Studies  - Glycogen Assay Kit (ab65620)
      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.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (46)

    Publishing research using ab65620? Please let us know so that we can cite the reference in this datasheet.

    ab65620 has been referenced in 46 publications.

    • Wilfred BS  et al. Alterations in Peripheral Organs following Combined Hypoxemia and Hemorrhagic Shock in a Rat Model of Penetrating Ballistic-Like Brain Injury. J Neurotrauma 37:656-664 (2020). PubMed: 31595817
    • Bruls YM  et al. Carnitine supplementation improves metabolic flexibility and skeletal muscle acetylcarnitine formation in volunteers with impaired glucose tolerance: A randomised controlled trial. EBioMedicine 49:318-330 (2019). PubMed: 31676389
    • Borg ML  et al. Modified Ucn2 Peptide Acts as an Insulin Sensitizer in Skeletal Muscle of Obese Mice. Diabetes N/A:N/A (2019). PubMed: 31010957
    • Hsu BE  et al. Immature Low-Density Neutrophils Exhibit Metabolic Flexibility that Facilitates Breast Cancer Liver Metastasis. Cell Rep 27:3902-3915.e6 (2019). PubMed: 31242422
    • Holland J  et al. A Brain-Melanocortin-Vagus Axis Mediates Adipose Tissue Expansion Independently of Energy Intake. Cell Rep 27:2399-2410.e6 (2019). PubMed: 31116984
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 45 Abreviews or Q&A

    Question

    I'm growing mouse primary hepatocytes on a collagen gel, lysing the cells in the gel, then continuing with the assay (this may result in some collagen inevitably being transferred as sample). In my sample and control wells (without hydrolysis buffer) as soon as I add the developer all wells turn very bright red and then turn clear after only a few minutes.
    Do you have any ideas what may be happening and advice on how to prevent this?

    Read More

    Abcam community

    Verified customer

    Asked on May 21 2014

    Answer

    Cells within collagen or Matrigel matrix are not ideal since when you homogenize the samples in the matrix and boil them, the matrix or components of the matrix can come along with the sample. This can affect the assay. Without knowing exactly what goes into this matrix, it is difficult to say what the affect will be and what component is inhibitory. We have not tested this type of sample in the culture matrix. For samples with reducing substances ab169558 is a better choice.

    Read More

    Kevin Hanson

    Abcam Scientific Support

    Answered on May 21 2014

    Question

    I'm growing mouse primary hepatocytes on a collagen gel, lysing the cells in the gel, then continuing with the assay (this may result in some collagen inevitably being transferred as sample). In my sample and control wells (without hydrolysis buffer) as soon as I add the developer all wells turn very bright red and then turn clear after only a few minutes.
    Do you have any ideas what may be happening and advice on how to prevent this?

    Read More

    Abcam community

    Verified customer

    Asked on May 16 2014

    Answer

    If you are using something like Matrigel for culturing the primary hepatocytes, then it is essential to remove the gel and collect the cells before proceeding with the homogenization and boiling to extract the glycogen. Liver cells store glycogen and hence it is critical to optimize the volume of the sample used per well such that the values are within the linear range of the std. curve.

    Read More

    Kevin Hanson

    Abcam Scientific Support

    Answered on May 16 2014

    Question

    How stable is the fluorescent signal?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 02 2014

    Answer

    The fluorescent signal is quite stable for several hours when protected from light. However, we recommend measuring the plate immediately after incubation for the best results.

    Read More

    Abcam Scientific Support

    Answered on Apr 02 2014

    Question

    In comparing these 2 Glycogen assay kits (Glycogen assay kit and glycogen assay kit II) I see that Assay Kit II requires no boiling step to inactivate enzymes in the sample. but Glycogen assay kit (I) does require a boiling step. Is this a mistake or is there something different about the buffers in assay Kit II that makes boiling unncecessary.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 12 2014

    Answer

    ab65620, Glycogen Assay Kit, can detect glycogen 0.0004 to 2 mg/ml. This kit can be used in both colorimetric and fluorometric applications. In this assay, glucoamylase hydrolyzes the glycogen to glucose which is then specifically oxidized to produce a product that reacts with OxiRed probe to generate color (λmax= 570 nm) and fluorescence (Ex 535/Em 587).

    Since the fluorometric version is very sensitive, this improved kit uses glycogen extraction and removes all enzyme activity by boiling to reduce glycogenolysis and the samples can be stored after this step to be assayed later.


    ab169558, Glycogen Assay Kit II (Colorimetric), can detect down to around 4 µg/ml of Glycogen in samples. This kit is colorimetric only. This assay is suitable for measuring Glycogen levels in samples that contain reducing substances, which may interfere with the oxidase-based assays like that used in ab65620. In this assay, Glycogen is hydrolyzed into glucose, which is oxidized to form an intermediate that reduces a colorless Probe to a colored product with strong absorbance at 450 nm.

    This is faster protocol without any glycogen extraction but not as sensitive as the fluorometric version.

    Read More

    Jeremy Kasanov

    Abcam Scientific Support

    Answered on Mar 12 2014

    Question

    I'm trying to decide how to prepare my tissue samples for use with this glycogen assay kit. Protocol states that for 10mg tissue to suspend tissue in 200ul water. But then it states that for larger masses of tissue (a few hundred milligrams or more) there is a KOH extraction step. My sample is 100 - 150mg muscle tissue. Should I resuspend this tissue in a relatively greater amount of water (eg. 2-3ml) or should I use the KOH extraction method?

    Read More

    Abcam community

    Verified customer

    Asked on Jan 31 2014

    Answer

    For 100-150mg of a tough tissue like muscle, I would definitely recommend using the KOH method. You can also consult the literature for any other glycogen extraction method that may be preferable.

    Read More

    Jeremy Kasanov

    Abcam Scientific Support

    Answered on Jan 31 2014

    Question


    Inquiry: Hi, A project partner of ours is interested in measuring glycogen levels in cattle (tissue samples). They want to complete the sample collection side of the work and then submit samples to us for analysis.
    Will we need to account for the residual glucose concentration in cattle by running 2 separate samples per animal and use the hydrolysis enzyme mix on only one of the samples?
    In terms of kit components, are 96 well plates included with the kit or should they be externally sourced by us? If the latter, what type of plate is recommended?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 12 2013

    Answer



    I can confirm that glucose generates background readings. Since glucose is present in your sample, we strongly recommend to do a glucose control without the addition of hydrolysis enzyme to determine the level of glucose background in your sample. The glucose background can then be subtracted from glycogen readings.
    Since every sample can be very different in its glucose levels (even from the same animal taken at different time points), we recommend to have two measurements for every sample: One with hydrolysis enzyme to measure total glucose (background glucose plus glycogen) and one without hydrolysis enzyme to measure glucose background alone.

    Plates are not included in the kit. Please use plates according to your detection preference:

    Fluorescence: black wall plates with clear bottoms.

    Colorimetry: Clear plates.

    Read More

    Anja Hoffmann

    Abcam Scientific Support

    Answered on Nov 12 2013

    Question


    Inquiry: Hello, I used several times your kits for the determination of glycogen in tissues homogenized in water and got good results. Now I need to use Levamisol 10mM for homogenization and I wonder if there may be interference with this kit. thank you for your help

    Read More

    Abcam community

    Verified customer

    Asked on Apr 18 2013

    Answer

    Thank you for contacting us.

    This kit was optimized using water, and unfortunately we don't have any data on whether Levamisol might work. This would have to be tested.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Apr 18 2013

    Question


    Inquiry: Hi, I used your Glycogen Assay kit for several months. I need to change buffer for the homogenization because I need the same homogenate for other dosing. I would like to use PBS or metaphosphoric acid (0.5-2%) instead of H2O. Do you know if i will be able to use your kit without any interference Thank you for your help

    Read More

    Abcam community

    Verified customer

    Asked on Feb 12 2013

    Answer

    Thank you for contacting us.

    PBS can be used in place of water for homgenizing.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Feb 12 2013

    Question

    I want to assay glycogen content in the cell using abcam kit. I would like to know whether I can cell lysis buffer to lyse cells and whether lysis buffer  will affect the measurement result of glycogen. If i can use lysis buffer, can you let me know which kind of buffer is fine for this assay,  is ripa buffer ok? whether need add protein inhibitor cocktail?

    Read More

    Abcam community

    Verified customer

    Asked on Jan 30 2013

    Answer

    Are you inquiring about ab65620? If so, you will not need to use any lysis buffers with this product. For tissue or cell samples with this kit you will homogenize 10^6 cells or 10 mg tissue with 200 μl dH₂O on ice. Then boil the homogenates for 5 min to inactivate enzymes. Spin the boiled samples at 13000 rpm for 5 min to remove insoluble material; the supernatant is ready for assay.

    Read More

    Abcam Scientific Support

    Answered on Jan 30 2013

    Question

    Please advise the shipping conditions of ab65620. Is it 4C or -20C?
    Thanks in advance.
    Best Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 14 2012

    Answer

    Thank you for contacting us.
    The shipping temperature for this kit is +4C and for storage temperature please refer to the protocol.
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
    Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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    Abcam Scientific Support

    Answered on Dec 14 2012

    1-10 of 45 Abreviews or Q&A

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