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Glycogen Colorimetric/Fluorometric Assay Kit

based on 48 citations in multiple journalsGlycogen Colorimetric/Fluorometric Assay Kit485 5
Catalog #: K646

In stock

$475.00

Product Details

Cat # +Size K646-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The assay can detect glycogen 0.0004 to 2 mg/ml.
Features & Benefits • Simple procedure; takes ~ 40 minutes
• Fast and convenient
• Kit contains all necessary reagents for accurate measurement of Glycogen
Kit Components • Hydrolysis Buffer
• Development Buffer
• OxiRed Probe
• Hydrolysis Enzyme Mix
• Development Enzyme Mix
• Glycogen Standard (2.0 mg/ml)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

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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Where is the glycogen standard from? Is it chemically synthesized?
The glycogen standard is from an animal-derived source.
Is there any glucose in the glycogen standard?
There should not be any glucose in the glycogen standard. Glucose in the samples can generate background in this assay.
Please explain the mechanism of detection. Is it the same as the Chan and Exton paper (1976) titled "A rapid method for the determination of glycogen content and radioactivity in small quantities of tissue or isolated hepatocytes " ?
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 (OD 570 nm) and fluorescence (Ex/Em = 535/587 nm). The assay can detect glycogen 0.0004 to 2 mg/ml. Our assay is non-radioactive. We do not precipitate samples on filter paper to assay glycogen as was done in the Chan and Exton paper. Based on the sample type, different methods of glycogen extraction are recommended on the datasheet
What are the differences between K646 and K648?
K646-100: 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 (?max= 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. K648-100: can detect less than 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 oxidasebased assays. 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 a faster protocol without any glycogen extraction but not as sensitive as the fluorometric version of K646.
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
How can mg of glycogen be converted to Molar concentration? How else can the data be reported?
mg of glycogen can be converted to mmoles of glycogen using molecular weight of glycogen (since the exact weight of glycogen in a sample can vary, it is a more precise way to report mg of glycogen). Instead of the per ml glycogen, mg of tissue used per well can be calculated based on the starting amount and the volume added to each well. In this way then mg of glycogen /mg tissue can be reported.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
What would be a good positive control sample tissue for glygogen in rat?
Liver is the main site for glycogen storage. Hence rat liver was used for our data on the datasheet and you can use this too.
Qin J et al., (2017) Effect of mild hypothermia preconditioning against low temperature (4°C) induced rat liver cell injury in vitro, PLoS One, 2017, 12(4): epub
Gu Z (2017) NEK2 Promotes Aerobic Glycolysis in Multiple Myeloma Through Regulating Splicing of Pyruvate Kinase, J Hematol Oncol
Ito, Tomohiro et al. (2017) OM-X®, a Fermented Vegetables Extract, Facilitates Muscle Endurance Capacity in Swimming Exercise Mice , Natural Product Communications. 2017;12(1):111-114
Wharfe, Michaela D. et al. (2016) Pregnancy-induced changes in the circadian expression of hepatic clock genes: implications for maternal glucose homeostasis, Am J Physiol Endocrinol Metab. 2016 Sep 1;311(3):E575-86.
Yang, Xiaodong et al. (2016) Physiological Expression of AMPKγ2RG Mutation Causes Wolff-Parkinson-White Syndrome and Induces Kidney Injury in Mice, J Biol Chem. 2016 Nov 4;291(45):23428-23439.
Fleet et al., SRC-2 orchestrates polygenic inputs for fine-tuning glucose homeostasis. PNAS, Nov 2015; 112: E6068 - E6077.
For more citations of this product click here