Product Details
| abID | ab102532 |
|---|---|
| Cat # +Size | K786-100 |
| Size | 100 assays |
| Detection Method | Absorbance (450 nm) |
| Species Reactivity | Mammalian |
| Applications | This assay detects GDH activity as low as 0.01 mU with our unit definition. |
| Features & Benefits | • Sensitive assays for measuring Glucose Dehydrogenase in various biological samples |
| Kit Components | • GDH Assay Buffer • Glucose (2 M) • Developer (Lyophilized) • GDH Positive Control (Lyophilized) • NADH Standard (0.5 μmol, Lyophilized) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Glucose 1-dehydrogenase (NADᶧ) (EC 1.1.1.118 ) is an enzyme that catalyzes the chemical reaction: D-glucose + NADᶧ↔D-glucono-1,5-lactone + NADH + H+. This enzyme belongs to the family of oxidoreductases, specifically those acting on the CH-OH group of donor with NADᶧ or NADPᶧ as acceptor. BioVision’s Glucose Dehydrogenase (GDH) Assay Kit provides a convenient tool for sensitive detection of the GDH in a variety of samples. The GDH present in sample will recognize D-glucose as a specific substrate leading to a proportional color development. The activity of GDH can be easily quantified colorimetrically (λ = 450 nm). This assay detects GDH activity as low as 0.01 mU with our unit definition.
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Does Glucose dehydrogenase catalyze reaction in both directions, foward and reverse ?
You are absolutely right in saying that the Glucose Dehydrogenase catalyzes the chemical reaction both ways, but for our particular assay, since we are providing glucose as the substrate and colour development as the end point, we detect only one sided Glucose Dehydrogenase reaction.
Can we use media with these kits? If so, how do we prepare it?
Yes, you can use cell culture media with these kits. Media would be one of the easiest samples preparation wise for these assays. There is no homogenization or pre-processing involved. Just aspirate out the media, and use it. Wherever mentioned on the datasheet, the media may need to be deproteinated before use. Use different volumes for the pilot expt to test out the optimal vol giving you a reading within the linear range of the standard curve.
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 yopur 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 -20C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are my 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 do I normalize my samples against protein concentration?
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without 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.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you 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.
Kamonwan Chamchoy, Application of WST-8 based colorimetric NAD(P)H detection for quantitative dehydrogenase assays. BMC Biochem, Apr 19; 30961528.
For more citations of this product click here
