Overview

  • Product name
    Glycogen Assay Kit
    See all Glycogen kits
  • Detection method
    Colorimetric/Fluorometric
  • Sample type
    Cell culture supernatant, Urine, Serum, Plasma, 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.

  • Platform
    Microplate reader

Properties

Images

  • 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.

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

  • Measurement of glycogen in various mouse tissues using 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

References

This product has been referenced in:
  • Lundell LS  et al. Regulation of glucose uptake and inflammation markers by FOXO1 and FOXO3 in skeletal muscle. Mol Metab 20:79-88 (2019). Read more (PubMed: 30502001) »
  • de Castro Barbosa T  et al. Paternal high-fat diet transgenerationally impacts hepatic immunometabolism. FASEB J 33:6269-6280 (2019). Read more (PubMed: 30768368) »
See all 34 Publications for this product

Customer reviews and Q&As

1-10 of 45 Abreviews or Q&A

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.

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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.

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Question
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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.
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1-10 of 45 Abreviews or Q&A

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