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Glycogen Colorimetric/Fluorometric Assay Kit Kit

BCA Reactivity: Chemical Fluorometric, Colorimetric Adherent Cell Culture, Cell Culture Cells, Cell Samples, Tissue Samples
Pubmed (38)
Catalog No. ABIN411676
$525.00
Plus shipping costs $45.00
100 tests
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  • Target See all Glycogen Kits
    Glycogen
    Reactivity
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    • 1
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    Chemical
    Detection Method
    Fluorometric, Colorimetric
    Detection Range
    00004-2 mg/mL
    Minimum Detection Limit
    0.0004 mg/mL
    Application
    Biochemical Assay (BCA)
    Sample Type
    Adherent Cell Culture, Cell Culture Cells, Cell Samples, Tissue Samples
    Specificity
    The Kit is an easy and convenient assay to measure glycogen levels in biological samples. In the assay, glucoamylase hydrolyzes the glycogen to glucose which is then specifically oxidized to produce a product that reacts with OxiRed probe to generate color (lambda max = 570 nm) and fluorescence (Ex 535/Em 587). The assay can detect glycogen 0.0004 to 2?mg/mL.
    Characteristics
    Glycogen Assay Kit: Colorimetric & Fluorometric Assay for measuring Glycogen levels in a variety of Samples such as Tissue, Cells etc. within 30 min. Simple & Convenient.
    Components
    Hydrolysis Buffer
    Development Buffer
    OxiRed Probe
    Hydrolysis Enzyme Mix
    Development Enzyme Mix
    Glycogen Standard (2.0 mg/ml)
  • Application Notes
    The assay can detect glycogen 0.0004 to 2 mg/mL.
    Comment

    Further details regarding sample type:

    • Animal tissues: Liver, muscle etc.
    • Cell culture: Adherent or suspension cells.

    Assay Time
    0.5 h
    Protocol
    1. Standard Curve Preparations: Colorimetric Assay: Dilute the Glycogen Standard to 0.2 mg /mL by adding 10 µL of the Standard to 90 µL of distilled water, mix well. Add 0, 2, 4, 6, 8, 10 µL to a series of wells. Adjust volume to 50 µL/well with Hydrolysis Buffer to generate 0, 0.4, 0.8, 1.2, 1.6 and 2.0 µg per well of the Glycogen Standard. Fluorometric Assay: Dilute the Glycogen Standard to 0.02 mg/mL by adding 10 µL of the Standard to 990 µL of distilled water, mix well. Add 0, 2, 4, 6, 8, 10 µL to a series of wells. Adjust volume to 50 µL/well with Hydrolysis Buffer to generate 0, 0.04, 0.08, 0.12, 0.16 and 0.2 µg per well of the Glycogen Standard.
    2. Sample Preparation*: Liquid samples can be assayed directly. For tissue or cells, homogenize 10^6 cells or 10 mg tissue with 200 µL dH2O on ice. Boiling the homogenates for 5 min to inactivate enzymes. Spin the boiled samples at 13000rpm for 5 min to remove insoluble material, the supernatant is ready for assay. Add up to 50 µL of sample or buffer (blank) to test wells. Adjust the volume to 50 µL with Hydrolysis Buffer. For unknown samples, we suggest testing several doses of your sample to ensure the readings are within the standard curve. * Notes: A. Glycogen can be metabolized very rapidly in some tissues after death (within a minute). Therefore, special cares must be taken to minimize glycogen consumption when take tissue sample, such as, frozen samples immediately, keep cold while working. B. There are a variety of methods for extraction of glycogen from tissues 1-4 depending upon the type of tissue or type of information desired, such as using 30 %KOH extraction/ethanol precipitation method to remove glucose background 5, or measure glycogen molecular weight distrubution 1, etc.
    3. Hydrolysis**: Colorimetric Fluorometric Hydrolysis Enzyme Mix 2 µL 1 µL Add Hydrolysis Enzyme Mix into Standard and samples, mix well, incubate for 30 minutes at room temperature. **Note: Glucose generates background readings (such as, culture medium, ect.). If glucose is present in your sample, you may 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.
    4. Development: Mix enough reagents for the number of samples and standards to be performed: For each well, prepare a total 50 µL Reaction Mix. Colorimetric Fluorometric Development Buffer 46 µL 48.7 µL Development Enzyme Mix 2 µL 1.0 µL OxiRed Probe 2 µL 0.3 µL Add 50 µL of the Reaction Mix to each well containing Glycogen Standard or samples.
    5. Incubate at room temperature for 30 minutes, protect from light.
    6. Measure colorimetrically (OD at 570 nm) or fluorometrically (Ex/Em 535/587 nm).
    Restrictions
    For Research Use only
  • Storage
    -20 °C
    Expiry Date
    12 months
  • Kramer, Grindley, Crowell, Makaron, Kohli, Kirby, Mansfield, Wachtman: "The common marmoset as a model for the study of nonalcoholic fatty liver disease and nonalcoholic steatohepatitis." in: Veterinary pathology, Vol. 52, Issue 2, pp. 404-13, 2015 (PubMed).

    Yang, Yu, Zhang, Tian, Qi, Wang: "Octopamine mediates starvation-induced hyperactivity in adult Drosophila." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 112, Issue 16, pp. 5219-24, 2015 (PubMed).

    Cansby, Nerstedt, Amrutkar, Durán, Smith, Mahlapuu: "Partial hepatic resistance to IL-6-induced inflammation develops in type 2 diabetic mice, while the anti-inflammatory effect of AMPK is maintained." in: Molecular and cellular endocrinology, Vol. 393, Issue 1-2, pp. 143-51, 2014 (PubMed).

    Fritz, Dong, Apsley, Martin, Na, Sitaraman, Weaver: "Deficiency of the BiP cochaperone ERdj4 causes constitutive endoplasmic reticulum stress and metabolic defects." in: Molecular biology of the cell, Vol. 25, Issue 4, pp. 431-40, 2014 (PubMed).

    Thatcher, Tippetts, Nelson, Swensen, Winden, Hansen, Anderson, Johnson, Porter, Reynolds, Bikman: "Ceramides mediate cigarette smoke-induced metabolic disruption in mice." in: American journal of physiology. Endocrinology and metabolism, Vol. 307, Issue 10, pp. E919-27, 2014 (PubMed).

    Witney, Carroll, Alam, Chandrashekran, Nguyen, Sala, Harris, DeBerardinis, Agarwal, Aboagye: "A novel radiotracer to image glycogen metabolism in tumors by positron emission tomography." in: Cancer research, Vol. 74, Issue 5, pp. 1319-28, 2014 (PubMed).

    Biesemann, Mendler, Wietelmann, Hermann, Schäfers, Krüger, Boettger, Borchardt, Braun: "Myostatin regulates energy homeostasis in the heart and prevents heart failure." in: Circulation research, Vol. 115, Issue 2, pp. 296-310, 2014 (PubMed).

    Lottes, Newton, Spyropoulos, Baatz: "Alveolar type II cells maintain bioenergetic homeostasis in hypoxia through metabolic and molecular adaptation." in: American journal of physiology. Lung cellular and molecular physiology, Vol. 306, Issue 10, pp. L947-55, 2014 (PubMed).

    Ong, Lee, Theodorou, Kodo, Lim, Shukla, Briston, Kiriakidis, Ashcroft, Davidson, Maxwell, Yellon, Hausenloy: "HIF-1 reduces ischaemia-reperfusion injury in the heart by targeting the mitochondrial permeability transition pore." in: Cardiovascular research, Vol. 104, Issue 1, pp. 24-36, 2014 (PubMed).

    Zager, Johnson, Becker: "Renal cortical pyruvate depletion during AKI." in: Journal of the American Society of Nephrology : JASN, Vol. 25, Issue 5, pp. 998-1012, 2014 (PubMed).

    Smith, Stallons, Schnellmann: "Renal cortical hexokinase and pentose phosphate pathway activation through the EGFR/Akt signaling pathway in endotoxin-induced acute kidney injury." in: American journal of physiology. Renal physiology, Vol. 307, Issue 4, pp. F435-44, 2014 (PubMed).

    Loffler, Birkenfeld, Philbrick, Belman, Habtemichael, Booth, Castorena, Choi, Jornayvaz, Gassaway, Lee, Cartee, Philbrick, Shulman, Samuel, Bogan: "Enhanced Fasting Glucose Turnover in Mice with Disrupted Action of TUG Protein in Skeletal Muscle." in: The Journal of biological chemistry, 2013 (PubMed).

    Hsu, Huang, Chen, Yen, Wu, Uang, Yang, Liu: "Antidiabetic effects of pterosin A, a small-molecular-weight natural product, on diabetic mouse models." in: Diabetes, Vol. 62, Issue 2, pp. 628-38, 2013 (PubMed).

    Wang, Sreenivasan, Gong, OConnell, Dabkowski, Hecker, Ionica, Konig, Mahurkar, Sun, Stanley, Sztalryd: "Cardiomyocyte-specific perilipin 5 overexpression leads to myocardial steatosis and modest cardiac dysfunction." in: Journal of lipid research, Vol. 54, Issue 4, pp. 953-65, 2013 (PubMed).

    Pearen, Goode, Fitzsimmons, Eriksson, Thomas, Cowin, Wang, Tuong, Muscat: "Transgenic muscle-specific Nor-1 expression regulates multiple pathways that effect adiposity, metabolism, and endurance." in: Molecular endocrinology (Baltimore, Md.), Vol. 27, Issue 11, pp. 1897-917, 2013 (PubMed).

    Solberg, Robertson, Aronsen, Rognmo, Sjaastad, Wisløff, Klungland: "Deletion of mouse Alkbh7 leads to obesity." in: Journal of molecular cell biology, Vol. 5, Issue 3, pp. 194-203, 2013 (PubMed).

    Hsieh, Lan, Liu, Zhang, Lin, Chen, Chiou: "Hepatitis C virus E2 protein involve in insulin resistance through an impairment of Akt/PKB and GSK3β signaling in hepatocytes." in: BMC gastroenterology, Vol. 12, pp. 74, 2012 (PubMed).

    Monserrate, Chen, Brachmann: "Drosophila larvae lacking the bcl-2 gene, buffy, are sensitive to nutrient stress, maintain increased basal target of rapamycin (Tor) signaling and exhibit characteristics of altered basal energy metabolism." in: BMC biology, Vol. 10, pp. 63, 2012 (PubMed).

    Drager, Li, Shin, Reinke, Aggarwal, Jun, Bevans-Fonti, Sztalryd, OByrne, Kroupa, Olivecrona, Blaner, Polotsky: "Intermittent hypoxia inhibits clearance of triglyceride-rich lipoproteins and inactivates adipose lipoprotein lipase in a mouse model of sleep apnoea." in: European heart journal, Vol. 33, Issue 6, pp. 783-90, 2012 (PubMed).

    German, Thaler, Wisse, Oh-I, Sarruf, Matsen, Fischer, Taborsky, Schwartz, Morton: "Leptin activates a novel CNS mechanism for insulin-independent normalization of severe diabetic hyperglycemia." in: Endocrinology, Vol. 152, Issue 2, pp. 394-404, 2011 (PubMed).

  • Target
    Glycogen
    Abstract
    Products
    Target Type
    Chemical
    Background
    Glycogen is the primary short term energy storage molecule in animals, synthesized primarily in the liver and muscle. Glycogen is a branched glucose polymer, in α-1,4 linkage, with branching via α-1,6 linkage. Abnormal ability to utilize glycogen is found in diabetes and in several genetic glycogen storage diseases. The BioVision Kit is an easy and accurate assay to measure glycogen levels in biological samples. In the assay, 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.
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