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Product Details
| Cat # +Size | K616-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm) |
| Species Reactivity | Mammalian |
| Applications | N/A |
| Features & Benefits | • Simple procedure; takes ~40 minutes • Fast and convenient • Kit provides all necessary buffers and reagents for a accurate assay of glucose and sucrose individually in various biological samples |
| Kit Components | • Glucose Assay Buffer • Glucose Probe (in DMSO) • Invertase (Lyophilized) • Glucose Enzyme Mix (Lyophilized) • Sucrose Standard (100 nmol/l) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Glucose (C₆H₁₂O₆; FW: 180.16) and sucrose (C₁₂H₂₂O₁₁; FW:342.3) are the important fuel sources to generate universal energy molecule ATP. Measurement of glucose or sucrose level can be very important in both research and development process. Sucrose is a disaccharide which can be converted into one glucose and one fructose when adding Invertase. BioVision’s Glucose and Sucrose Assay Kit provides a convenient means for measuring glucose and sucrose levels from various biological samples (e.g. serum, plasma, body fluids, food, growth medium, etc.). To measure glucose level, glucose oxidase specifically oxidizes free-glucose generating a compound that reacts with the glucose probe to produce resorufin, which can be detected colorimetrically (O.D. 570 nm) or fluorometrically (Ex/Em 535/587). To measure sucrose, invertase can be added to the reaction to convert sucrose to free glucose and fructose, so total glucose level can be measured. Then the sucrose level = Total Glucose – Free Glucose.
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Should a glucose standard be prepared since the kit comes only with a Sucrose standard?
This is not necessary since the invertase converts sucrose to glucose and fructose. This glucose is measured in the reaction with the probe. A separate std curve with glucose will yield identical results.
Will disaccharides other than sucrose be measured by this kit?
The data on the datasheet shows that other disaccharides like Lactose and Maltose do not interfere in this kit's measurements. Nedogenous glucose can generate background and needs to be subtracted using a background control.
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.
Can we use frozen samples 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 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 exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
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 and 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.
Why are the standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How are samples normalized against protein concentration?
A protein quantitation assay can be used with the supernatants from cell/tissue lysates or with any other liquid sample in the assay buffer.
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 we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
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.
