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MAK016
Glycogen Assay Kit
sufficient for 100 colorimetric or fluorometric tests
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About This Item
UNSPSC Code:
12161503
NACRES:
NA.84
Recommended Products
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
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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Danger
Hazard Statements
Precautionary Statements
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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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
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)
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
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
Davide Ghinolfi et al.
Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society, 25(3), 436-449 (2018-10-27)
Ex situ normothermic machine perfusion (NMP) might minimize ischemia/reperfusion injury (IRI) of liver grafts. In this study, 20 primary liver transplantation recipients of older grafts (≥70 years) were randomized 1:1 to NMP or cold storage (CS) groups. The primary study endpoint
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What is the polymer size of glycogen used in the standard of the MAK016-1KT Glycogen Assay Kit?
1 answer-
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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-
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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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-
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.
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Is it possible to know information about compatibility with extreme pH, rather than with salt buffers etc...?
1 answer-
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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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-
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.pdfHelpful?
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