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Total Carbohydrate Colorimetric Assay Kit

based on 1 citations in multiple journalsTotal Carbohydrate Colorimetric Assay Kit14.1 4
Robust, Sensitive Assay
Catalog #: K645
$490.00

Product Details

Cat # +Size K645-100
Size 100 assays
Detection Method Absorbance (490 nm)
Applications Can detect most forms of carbohydrates, including simple & complex saccharides, glycans, glycoproteins & glycolipids
Features & Benefits • Simple procedure; takes ~ 30 minutes
• Rapid, convenient and sensitive
• Kit contains all necessary reagents for measuring total carbohydrate concentration
Kit Components • Assay Buffer
• Developer
• Standard (D-Glucose, 2 mg/ml)
Storage Conditions 4°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Carbohydrates play important structural as well as chemical roles in all living systems. Detection of total carbohydrates, therefore, has wide applications. BioVision’s Total Carbohydrate Assay is a simple, sensitive and robust method of detecting virtually all carbohydrates. The assay is based on the phenol-sulfuric acid method. In Total Carbohydrate Assay, polysaccharides (mono, di, tri, etc.) and their derivatives, in the presence of sulfuric acid, are hydrolyzed to monomers and converted to furfural or hydroxyfurfural, which react with the Developer to form a chromogen that can be quantified by measuring the absorbance at 490 nm. The Total Carbohydrate Assay can detect most forms of carbohydrates, including simple and complex saccharides, glycans, glycoproteins and glycolipids.


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Will the Total Carbohydrate Colorimetric Assay Kit identify N-acetylglucosamine and N-acetylchitobiose, which are the expected degradation products of Chitin?
The glycosidic linkages of Chitosan are resistant to acid hydrolysis such as the phenol-sulphuric acid method, which is used in K645. Hence we do not expect determination of Chitin using K645.
Have you ever done measurements to study interference from different oxidising substances? We have used the Anthrone method for carbohydrate analysis, however, the presence of nitrate and nitrite interfered with the analysis severely which is why we are concerned.
The assay buffer contains NP-40 and the developer contains phenol. For both of these, strong oxidizing agents should be avoided in the assay. We have not tested Nitrate/nitrite with this assay to check for any possible interference.
Will this kit quantify total carbohydrate or total soluble carbohydrate?
We have not distinguished between soluble and insoluble carbohydrates while testing this kit. All carbohydrates should be hydrolyzed with sulfuric acid. So this kit is to quantify total carbohydrates in the sample. 4. Simple sugars, oligosaccharides, polysaccharides and their derivatives can be quantified using this kit.
Is the principle of this kit same as phenol-sulfuric acid reaction reported by M.Duvois et al. (1956)?
Yes. The Dubois paper in 1956 demonstrates how phenol and sulfuric acid can provide a way to colorimetrically determine amount of carbohydrates (sugars and their methyl derivatives, oligosaccharides, and polysaccharides) in a sample.
How can the sensitivity be increased? For example what is the incubation time, temperature, how to add H2SO4 into the sample etc.
The conditions in the assay datasheet are good for a variety of samples including human serum, Jurkat cells, food components etc. The acid hydrolysis step can be optimized by the user but the 150ul acid, 15 mins at 90C has worked for us and most of our customers. Adding the sulfuric acid does not require any special procedure. Simply add 150 µl concentrated H2SO4 to the well, shake and incubate at 90°C for 15 min. The acid can be added inside a chemical hood for protective purposes.
As peptidoglycan is a polymer consisting of sugars and amino acids, this kit should work, shouldn’t it?
We have not tested this kit with peptidoglycan. The glycan portion theoretically should be detected by the kit. The acid hydrolysis might not break down all the carbohydrate in the peptidoglycan polymer to yield detectable furfural.
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.
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.
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.
What is the exact volume of sample required for this assay?
This depends on the sample and has to be optimized for each sample type by the user for best results.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted.
How important is the temperature incubation (90ºC) in the 3rd step of the protocol? Could it be done with less temperature but more time? Is 70ºC for 20-25 min acceptable?
Heating to 90°C is a very important step for the reaction. If you do not have a heating block that can get to 90°C, you can boil water and keep your samples in the boiling water for 15 min.
Karav, Sercan et al. (2016) Oligosaccharides Released from Milk Glycoproteins Are Selective Growth Substrates for Infant-Associated Bifidobacteria Appl Environ Microbiol. 2016 May 31;82(12):3622-30.
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