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D-Sorbitol Colorimetric Assay Kit

based on 2 citations in multiple journalsD-Sorbitol Colorimetric Assay Kit24.1 4
Catalog #: K631
$490.00

Product Details

Cat # +Size K631-100
Size 100 assays
Detection Method Absorbance (560 nm)
Applications The assay is useful over the range of 0.1-10 nmol of Sorbitol per sample.
Features & Benefits • Simple procedure; takes ~ 35-40 minutes
• Fast and convenient
• The kit provides an accurate assay for measuring sorbitol in various samples
Kit Components • Sorbitol Assay Buffer
• Sorbitol Probe
• Sorbitol Enzyme Mix
• Sorbitol Developer
• Sorbitol Standard (100 mM)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Sorbitol is one of the 6 carbon sugar alcohols. It is commonly used as an artificial sweetener, as a laxative and in cosmetics as a humectant and thickening agent. Sorbitol is produced naturally in a variety of fruits. It can be produced in humans in small amounts by the reduction of glucose by aldose reductase. Due to its poor ability to diffuse across the cell membrane, sorbitol can be trapped in cells and is believed to be one of the causes of damage (due to osmotic effects) in diabetes. Interestingly, sorbitol can be used as a screen for the O154:H7 strain of E. coli, since this strain is one of the few strains which cannot metabolize sorbitol. BioVision’s Sorbitol Colorimetric Assay Kit is designed to measure sorbitol in a variety of samples such as foods, fruits, fruit juices, pharmaceuticals, cosmetics, paper and some other biological samples. In the assay, sorbitol is oxidized to fructose with the proportional development of intense color with an absorbance maximum at 560 nm. The assay is useful over the range of 0.1-10 nmol of Sorbitol per sample.


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What is the product being measured by this kit?
Fructose is formed in the first step of the detection reaction and concomittantly the probe and the developer react with the byproduct of this reaction form a colored compound which can be measured at 560nm. Fructose formation is proportional to Sorbitol.
While using cell lysates from cells, the background was higher without the enzyme than with the enzyme. Would it be possible that the enzyme converts forwards as well as backwards?
The enzyme Sorbitol dehydrogenase catalyzes reversible reactions between Sorbitol and Fructose. Hemoglobin and/or Bilirubin can produce interference in this assay. If the cells are hematopoietic lineage cells, this can be one possibility.
If the detection enzyme catalyses the reaction bidirectionally, what makes sure that Sorbitol to Fructose catalysis will happen in the assay and not the other way around?
Yes Sorbitol dehydrogenase does catalyze reaction in both directions . The direction is decided by cofactor availability for the forward reaction. The cofactor is provided by the reaction mix components. If the backward reaction is predominant, there would be no color development.
Can this kit be used for measuring Sorbitol in RBC?
Since there is hemoglobin in RBS which can interfere with this assay, RBC is not a compatible sample type. This asay is also not recommended to be used with plasma, serum or urine.
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.
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.
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.
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 cells/tissues 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 we use the kit for cell culture or cell lysate samples?
Yes, the kit can be used for cell culture or cell lysate samples. Samples with unknown quantities of sorbitol should be run at varying dilutions to ensure that the readings fall within the linear portion of the standard curve.
Andres-Hernando, Ana et al. (2017) Protective role of fructokinase blockade in the pathogenesis of acute kidney injury in mice, Nat Commun. 2017 Feb 13;8:14181.
Lanaspa et al., Endogenous Fructose Production and Fructokinase Activation Mediate Renal Injury in Diabetic Nephropathy.J. Am. Soc. Nephrol., Nov 2014; 25: 2526 - 2538.
For more citations of this product click here