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Product Details
| Cat # +Size | K687-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Fluorescence (Ex/Em 535/587 nm) |
| Species Reactivity | Mammalian |
| Applications | The kit can detect G6P in the range of 10 to 500 pmoles which is equivalent to the range of 1-500 µM in the original sample, in a high-throughput screening (HTS) format. |
| Features & Benefits | • Simple procedure; takes ~ 35-40 minutes to complete the assay • Fast and convenient • Sensitive, efficient and accurate assay |
| Kit Components | • G6P Assay Buffer • PicoProbe™ (in DMSO) • G6P Enzyme Mix • G6P Substrate Mix • G6P Standard (10 µmol) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Glucose-6-phosphate (G6P) is a key intermediate for glucose transport into cells which then enters either metabolic pathways or storage. G6P can enter the glycolytic pathway, the pentose phosphate shunt or be stored as glycogen or starch. G6P is utilized by its dehydrogenase to generate reducing equivalents in the form of NAD(P)H. This is particularly important in red blood cells where a G6PDH deficiency leads to hemolytic anemia. BioVision's glucose-6-phosphate Assay Kit is a simple, sensitive and rapid means of quantifying G6P in a variety of samples. In the assay, glucose-6-phosphate is oxidized with the generation of a product that converts a nearly colorless probe to an intensely fluorescent product (Ex/Em 535/587 nm). The Glucose-6-phosphate Assay Kit can detect G6P in the range of 10 to 500 pmoles which is equivalent to the range of 1-500 µM in the original sample assuming a dilution of 5X during processing. The PicoProbe™ G6P Assay Kit is much more sensitive than the G6P Colorimetric Assay Kit for low G6P samples.
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Will samples in RIPA buffer work with this kit?
We recommend our assay buffer for tissue homogenization. RIPA buffer typically contains SDS and this is why we recommend avoiding its use. There are enzymes in the kit whose activity might be affected adversely by this.
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 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. It is recommended 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.
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.
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, the incubation time can be increased. As long as the standard curve is linear, it should be fine to use, since all the 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 alternate buffers be used 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.
Tan et al., The Monocarboxylate Transporter 4 Is Required for Glycolytic Reprogramming and Inflammatory Response in Macrophages. J. Biol. Chem., Jan 2015; 290: 46 - 55.
For more citations of this product click here
