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Glycogen assay kit

Manufactured by Cayman Chemical
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The Glycogen Assay Kit is a laboratory tool used to quantify the amount of glycogen present in samples. It provides a colorimetric method for the determination of glycogen levels.

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Close spelling variants of this product

Glycogen Fluorometric Assay Kit

22 protocols using glycogen assay kit

1

Quantifying Tissue Glycogen Levels

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Glycogen concentration was measured using Glycogen Assay Kit (Cayman Chemical, Ann Arbor, MI), according to the manufacturer’s instructions. Briefly, the frozen tissues were minced into small pieces, and then homogenized to disrupt cells. Homogenate was centrifuged and 50 μl of supernatant was used for glycogen measurement. The reagents of Glycogen Assay Kit (Cayman Chemical) were able to hydrolyze glycogen and generate highly fluorescent product resorufin (Glycogen Assay Kit Manual). Fluorescence (Ex/Em = 535/587 nm) was measured with the SpectraMax M3 system (Molecular Devices, Sunnyvale, CA).
Liang G., Malmuthuge N., Guan Y., Ren Y., Griebel P.J, & Guan L.L. (2016). Altered microRNA expression and pre-mRNA splicing events reveal new mechanisms associated with early stage Mycobacterium avium subspecies paratuberculosis infection. Scientific Reports, 6, 24964.
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2

Glycogen Quantification in Mouse Tissues

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After a 4 h fast the mice were sacrificed with CO2 and 150 mg tissue samples were collected from the superior lobe of the liver, the placenta, kidney, and skeletal muscle (M. quadriceps femoris). These samples were snap frozen in liquid nitrogen and stored in −80°C before analysis with a fluorometric Glycogen Assay Kit (700480, Cayman Chemical).
Sissala N., Myllymäki E., Mohr F., Halmetoja R., Kuvaja P., Dimova E.Y, & Koivunen P. (2022). Hypoxia ameliorates maternal diet‐induced insulin resistance during pregnancy while having a detrimental effect on the placenta. Physiological Reports, 10(9), e15302.
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3

Cotton Aphid Glycogen and Glucose-6-Phosphate Assay

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Samples of 2–3 instar cotton aphids were obtained from cotton aphids after being parasitized for 8 h, 16 h, 1 d, 2 d, and 3 d. Samples of non-parasitized cotton aphids were obtained as controls (the larvae of wasps were removed under the microscope, and the control samples were also dissected). The contents of the glycogen and 6-phosphate glucose contents in cotton aphids parasitized for different periods of time, as well as the control, were detected using reagent kits (Glycogen Assay Kit and Glucose-6-Phosphate (G6P) Fluorometric Assay Kit, Cayman Chemical, Ann Arbor, MI, USA). SPSS 20.0 was used to evaluate the differences among copy numbers with Mann-Whitney U test (n = 2) and KruskaleWallis test (n > 2).
Gao X., Xue H., Luo J., Ji J., Zhang L., Niu L., Zhu X., Wang L., Zhang S, & Cui J. (2020). Molecular Evidence that Lysiphlebia japonica Regulates the Development and Physiological Metabolism of Aphis gossypii. International Journal of Molecular Sciences, 21(13), 4610.
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4

Platelet Glucose Uptake and Metabolism

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Platelet and megakaryocyte glucose uptake analysis were conducted with 10mM 3H-2-Deoxy-D-Glucose. 13C-glucose flux was determined through incubations of platelets with 25mM 1,6-13C-glucose for up to one hour; 13C-lactic acid was quantified with gas chromatography-mass spectroscopy. Glycogen content was determined in 2×108 CD45 and Ter119 bead depleted platelets using a Glycogen Assay Kit (Cayman Chemical, Ann Arbor, MI). Detailed protocols can be found in the supplemental methods.
Fidler T.P., Campbell R.A., Funari T., Dunne N., Angeles E.B., Middleton E., Chaudhuri D., Weyrch A.S, & Abel E.D. (2017). Deletion of GLUT1 and GLUT3 Reveal Multiple Roles for Glucose Metabolism in Platelet and Megakaryocyte Function. Cell reports, 20(4), 881-894.
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5

Glycogen Quantification in Liver and Muscle

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Liver and gastrocnemius muscle from WT and MafAS64F/+ mice (n = 5 per genotype and sex) were collected and flash frozen in liquid nitrogen. Tissue was homogenized with a Dounce homogenizer prior to determining glycogen content using a glycogen assay kit (Cayman Chemical) according to the manufacturer’s protocol.
Walker E.M., Cha J., Tong X., Guo M., Liu J.H., Yu S., Iacovazzo D., Mauvais-Jarvis F., Flanagan S.E., Korbonits M., Stafford J., Jacobson D.A, & Stein R. (2021). Sex-biased islet β cell dysfunction is caused by the MODY MAFA S64F variant by inducing premature aging and senescence in males. Cell reports, 37(2), 109813.
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6

Glycogen Content Quantification

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Glycogen levels were evaluated using a Glycogen assay kit (Cayman, 700480, Ann Arbor, MI, USA).
Benzi A., Spinelli S., Sturla L., Heine M., Fischer A.W., Koch-Nolte F., Mittrücker H.W., Guse A.H., De Flora A., Heeren J, & Bruzzone S. (2022). Role of Liver CD38 in the Regulation of Metabolic Pathways during Cold-Induced Thermogenesis in Mice. Cells, 11(23), 3812.
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7

Serum Lipid and Glucose Analysis in Mice

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Mice were anesthetized with isoflurane and blood collected via the orbital vascular plexus. The blood was centrifuged and the serum collected to detect the free fatty acids and low-density lipoprotein (LDL) using a fatty acid quantitation kit and LDL quantitation kit (Sigma), respectively, according to the manufacturer’s protocol as described previously [26 (link)]. The serum levels of glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), total cholesterol (TC), high-density lipoprotein (HDL), and total TGs were measured using the biochemical analyzer (DRI-CHEM NX500, Fuji, Tokyo, Japan). The day before the end of the experiment, mice fasted for 16 h and were administered glucose by intraperitoneal injection to assay blood glucose levels using the biochemical analyzer (Fuji). Blood insulin was detected using the Insulin EIA Kit (Cayman, Ann Arbor, Michigan, USA) according to the manufacturer’s protocol. Liver glycogen was detected using the Glycogen Assay Kit (Cayman) according to the manufacturer’s protocol, and the glycogen levels were measured using a microplate reader (Multiskan FC) at an absorbance of 570 nm.
Liou C.J., Wu S.J., Shen S.C., Chen L.C., Chen Y.L, & Huang W.C. (2020). Phloretin ameliorates hepatic steatosis through regulation of lipogenesis and Sirt1/AMPK signaling in obese mice. Cell & Bioscience, 10, 114.
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8

Quantifying Glycogen in Skeletal Muscle

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Glycogen content in SKM was determined using a glycogen assay kit (Cayman Chemical), according to the manufacturer’s instructions.
Meister J., Bone D.B., Knudsen J.R., Barella L.F., Velenosi T.J., Akhmedov D., Lee R.J., Cohen A.H., Gavrilova O., Cui Y., Karsenty G., Chen M., Weinstein L.S., Kleinert M., Berdeaux R., Jensen T.E., Richter E.A, & Wess J. (2022). Clenbuterol exerts antidiabetic activity through metabolic reprogramming of skeletal muscle cells. Nature Communications, 13, 22.
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9

Measuring Cellular Metabolism in Activated T Cells

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Intracellular glycogen levels were measured using the Glycogen Assay Kit (700480, Cayman Chemical) according to the manufacturer’s instructions. To determine glucose uptake and lactate production, naïve CD4+ T cells were activated with anti-CD3 and anti-CD28 beads for 4 days, and beads were removed. After washing, 1 million cells were cultured in 1-ml fresh normal medium for 24 hours and lactate production was determined using the Lactate Colorimetric/Fluorometric Assay Kit (K607, BioVision). Glucose uptake in activated CD4+ T cells was determined by incubating cells in glucose-free medium containing 200 μM 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-d-glucose (2-NBDG) (11046, Cayman Chemical) for 2 hours at 37°C, followed by flow cytometry–based analysis.
Jin J., Li X., Hu B., Kim C., Cao W., Zhang H., Weyand C.M, & Goronzy J.J. (2020). FOXO1 deficiency impairs proteostasis in aged T cells. Science Advances, 6(17), eaba1808.
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10

Treadmill Exercise and Glucose Tolerance

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Mice were fasted for 4 h and run on a treadmill for 1 h, using the following protocol: 5 min at 10 m/min, 50 min at 15 m/min, and then 5 min at 18 m/min. At the end of the running protocol, a glucose tolerance test was performed. One week later the experiment was repeated, but mice were euthanized 15 min after the i.p. injection of glucose (2 g/kg). SKM tissues were isolated and snap frozen in liquid nitrogen for measurement of glycogen content and western blotting analysis. Glycogen levels were determined using a glycogen assay kit (Cayman Chemical).
Meister J., Bone D.B., Godlewski G., Liu Z., Lee R.J., Vishnivetskiy S.A., Gurevich V.V., Springer D., Kunos G, & Wess J. (2019). Metabolic effects of skeletal muscle-specific deletion of beta-arrestin-1 and -2 in mice. PLoS Genetics, 15(10), e1008424.
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