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Glycogen Colorimetric Assay Kit II

based on 5 citations in multiple journalsGlycogen Colorimetric Assay Kit II54.1 4
For samples having reducing substances
Catalog #: K648

In stock

$485.00

Product Details

Cat # +Size K648-100
Size 100 assays
Detection Method Absorbance (450 nm)
Applications The assay can detect less than 4 µg/ml of Glycogen in various tissues. Analysis of metabolism and cell signaling.
Features & Benefits • Simple, rapid, and high-throughput adaptable.
• Kit contains all necessary reagents for accurate measurement of Glycogen levels.
Kit Components • Glycogen Hydrolysis Buffer
• Gycogen Development Buffer
• Hydrolysis Enzyme Mix (lyophilized)
• Development Enzyme Mix (lyophilized)
• Probe (lyophilized)
• Glycogen Standard (2 mg/ml)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Glycogen serves as the main carbohydrate storage in animals and can be converted to glucose readily. It is primarily found in the liver and muscle tissues. Glycogen is a branched biopolymer comprising of α-1,4 linkage with α-1,6 linkages occurring every 8-10 glucose units along the backbone. Abnormal ability to utilize glycogen is found in diabetes and in several genetic glycogen storage diseases. Biovision’s Glycogen Assay kit II provides a simple, fast and robust way to measure Glycogen levels in various biological samples. This assay is suitable for measuring Glycogen levels in samples that contain reducing substances, which may interfere with the oxidase-based 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 high-throughput suitable assay kit can detect less than 4 µg/ml of Glycogen in samples.


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What are the differences between K646 and K648?
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.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).
How should different dilutions of the sample be tested with this kit? Should the volumes be different in the well for each dilution?
Samples can be diluted over a range, for example, 1:10 or 1:50 or 1:100 or as required in a separate tube and then add 5ul for all dilutions, 10ul for all dilutions and so on into the wells. Then fill up volume to 50ul. This can be tested with one or two samples to optimize the dilution and the volume needed. If there is glucose in the sample, this can create background. A parallel background control can be used to subtract this value.
Why is there only 48 ul reaction mix for samples and standards where as there is 50 ul for the background?
The standard and the samples have 2ul of the hydrolysis enzyme mix (step 3, section VIII). This is why there is 48ul total reaction mix for the standards and the samples where as there is 50ul reaction mix for the background control.
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.
Is it possible to use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
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 is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature (4C) and in appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Liu et al., Irisin inhibits hepatic gluconeogenesis and increases glycogen synthesis via the PI3K/Akt pathway in type 2 diabetic mice and hepatocytes.Clin. Sci., Aug 2015; 129: 839 - 850.
Shemesh et al., Suppression of mTORC1 activation in acid-α-glucosidase-deficient cells and mice is ameliorated by leucine supplementation. Am J Physiol Regulatory Integrative Comp Physiol, Nov 2014; 307: R1251 - R1259.
Wu et al., Hypothalamic Nesfatin-1/NUCB2 Knockdown Augments Hepatic Gluconeogenesis That Is Correlated With Inhibition of mTOR-STAT3 Signaling Pathway in Rats.Diabetes, Apr 2014; 63: 1234 - 1247.
Anja Solberg et al., Deletion of mouse Alkbh7 leads to obesity. J Mol Cell Biol, Jun 2013; 5: 194 - 203.
Horng-Yih Ou et al., Multiple mechanisms of GW-9508, a selective G protein-coupled receptor 40 agonist, in the regulation of glucose homeostasis and insulin sensitivity. Am J Physiol Endocrinol Metab, Mar 2013; 304: E668 - E676.
For more citations of this product click here