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MAK016

Sigma-Aldrich

Glycogen Assay Kit

sufficient for 100 colorimetric or fluorometric tests

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.84

usage

sufficient for 100 colorimetric or fluorometric tests

detection method

colorimetric
fluorometric

storage temp.

−20°C

General description

Glycogen is a branched polymer of glucose that serves as the primary short-term energy storage molecule in animals. Glycogen is primarily synthesized in liver and muscle tissue where it can constitute up to 10% of the weight of liver and 1-2% of the weight of muscle tissue. While muscle glycogen is generally utilized locally, liver glycogen serves as an important buffer to regulate blood glucose levels. Glycogen metabolism is dysregulated in diabetes and the glycogen storage diseases due to inborn errors of metabolism.

Application

Glycogen Assay Kit has been used in glycogen quantification.[1] It has also been used to determine the glycogen content in liver[2][3], vastus lateralis and muscle homogenates.[4]

Suitability

Suitable for determining glycogen concentration in various tissues such as liver etc. and cell culture (adherent or suspension cells).

Principle

Glycogen concentration is determined by a coupled enzyme assay, which produces a colorimetric (570 nm)/ fluorometric (λex = 535/λem = 587 nm) product, proportional to the glycogen present.

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Product No.
Description
Pricing

Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H317,H334

Hazard Classifications

Resp. Sens. 1 - Skin Sens. 1

Storage Class Code

10 - Combustible liquids

Flash Point(F)

188.6 °F - closed cup

Flash Point(C)

87 °C - closed cup

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Certificates of Analysis (COA)

Lot/Batch Number
8J19K06460
8J15K06460
8H18K06460
8E10K06460
8B02K06460

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Madhuri S Salker et al.
Scientific reports, 7(1), 12612-12612 (2017-10-05)
Embryo implantation requires a hospitable uterine environment. A key metabolic change that occurs during the peri-implantation period, and throughout early pregnancy, is the rise in endometrial glycogen content. Glycogen accumulation requires prior cellular uptake of glucose. Here we show that
Insulin Signaling in Bupivacaine-induced Cardiac ToxicitySensitization during Recovery and Potentiation by Lipid Emulsion
Fettiplace M R, et al.
Anesthesiology, 124(2), 428-\442-428-\442 (2016)
Cassius E O Coombs et al.
Meat science, 134, 86-97 (2017-08-05)
This study evaluated the effect of chilled followed by frozen storage on lamb quality and safety parameters. Experimental (n=360) M. longissimus lumborum (LL) were randomly sampled from the boning room of a commercial Australian abattoir, at 24 h post-mortem, and
Nady Golestaneh et al.
Journal of translational medicine, 14(1), 344-344 (2016-12-22)
Study of age related macular degeneration (AMD) has been hampered by lack of human models that represent the complexity of the disease. Here we have developed a human in vitro disease model of AMD to investigate the underlying AMD disease
Long-term obestatin treatment of mice type 2 diabetes increases insulin sensitivity and improves liver function.
Kolodziejski P A, et al.
Endocrine, 56(3), 538-550 (2017)
View All Related Papers

Questions

1–7 of 7 Questions  

  1. How is shipping temperature determined? And how is it related to the product storage temperature?

    1 answer

    1. Products may be shipped at a different temperature than the recommended long-term storage temperature. If the product quality is sensitive to short-term exposure to conditions other than the recommended long-term storage, it will be shipped on wet or dry-ice. If the product quality is NOT affected by short-term exposure to conditions other than the recommended long-term storage, it will be shipped at ambient temperature. As shipping routes are configured for minimum transit times, shipping at ambient temperature helps control shipping costs for our customers. For more information, please refer to the Storage and Transport Conditions document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/316/622/storage-transport-conditions-mk.pdf

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  2. How can I determine the shelf life / expiration / retest date of this product?

    1 answer

    1. If this product has an expiration or retest date, it will be shown on the Certificate of Analysis (COA, CofA). If there is no retest or expiration date listed on the product's COA, we do not have suitable stability data to determine a shelf life. For these products, the only date on the COA will be the release date; a retest, expiration, or use-by-date will not be displayed.
      For all products, we recommend handling per defined conditions as printed in our product literature and website product descriptions. We recommend that products should be routinely inspected by customers to ensure they perform as expected.
      For products without retest or expiration dates, our standard warranty of 1 year from the date of shipment is applicable.
      For more information, please refer to the Product Dating Information document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/449/386/product-dating-information-mk.pdf

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  3. What could be the reason for the varying cloudiness levels in dilutions of INS-1 beta cell lysate when using the glycogen assay kit (cat #MAK016)? The solution became cloudy after dilution, with different dilutions performed: 1:2, 1:5, and 1:10. The 1:2 dilution appeared cloudier than the 1:5 and 1:10 dilutions, and the 1:5 dilution was cloudier than the 1:10 dilution. The cell lysate was clear after centrifugation before adding it to the hydrolysis buffer.

    1 answer

    1. The observed precipitation may be due to the detergent in the assay buffer when added to the sample. To prevent this, it is recommended to bring the assay buffer to room temperature before adding it. Dilution has shown to reduce turbidity, so using the dilution with the least turbidity is advisable. Testing the performance of this dilution with the standard curve would be beneficial. Considering the high acidity of the hydrolysis buffer, it is uncertain if the pH changes upon adding the lysate, potentially causing precipitation. Testing the pH of the lysate using a pH strip and adjusting it to around 4.5 with HCl, if necessary, while keeping in mind the confirmed pH of the cell lysate is around 7, can be done.

      Helpful?

  4. Is it permissible to proceed with boiling the liver samples homogenized in NP40 Substitute assay regent, even though the protocol states that "Tissue (10 mg) can be homogenized in 100 uL of water on ice. Boil homogenates for 5 minutes to inactivate enzymes"?

    1 answer

    1. It is not certain whether the samples prepared in the NP40 Substitute assay reagent will be compatible with this kit. However, one can proceed to the next step by boiling the homogenates for 10 minutes to inactivate enzymes. Following that, the mixture should be centrifuged at 18000 rpm for 10 minutes to remove insoluble material. Collect the supernatant for the assay.

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  5. What is the polymer size of glycogen used in the standard of the MAK016-1KT Glycogen Assay Kit?

    1 answer

    1. This information is proprietary and cannot be disclosed.

      Helpful?

  6. Is it possible to know information about compatibility with extreme pH, rather than with salt buffers etc...?

    1 answer

    1. The kit is tested with cell culture supernatant, biological fluids, tissue, and urine. It has not been tested with a sample that has an extreme pH. It is not known how that would affect the assay performance, as it has not been tested. However, the assay does work for urine which can have a wide pH range from pH 5-8.

      In the protocol, glycogen is extracted using first KOH and then further extracted using HCl. The precipitate is then dissolved in Development Buffer/Hydrolysis Buffer for analysis which is buffered for the assay.

      If the pH is already very basic this may be fine as it could be adjusted to the correct pH but if it is very acidic then the extraction may be difficult since the first step is to raise the pH. The buffers in the kit are also pH dependent as well so downstream steps require a specific pH.

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  7. We are looking for a Colorimetric/Fluorometric assay to determine glycogen content in various mouse tissues samples. We came across this kit and would like to know how much sample is required for this assay (volume/well).

    1 answer

    1. The minimum tissue requirement is 10mg in 200 uL. After homogenization and cleanup, a range of 2-50 uL of the sample will be added to each well. For unknown samples it is advised to test several sample dilutions to ensure readings are within linear range. Please see the link below for the Product Information Sheet:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/157/372/mak016bul.pdf

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