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PicoProbe™ Dihydroxyacetone Phosphate (DHAP) Fluorometric Assay Kit

Only Kit on the market to measure DHAP
Catalog #: K673
$665.00

Product Details

Cat # +Size K673-100
Size 100 assays
Detection Method Fluorescence (Ex/Em = 535/587 nm)
Applications Measurement of Dhydroxyacetone Phosphate in various tissues/cells
Analysis of metabolic pathways such as glycolysis, Calvin cycle in plants, methylglyoxal pathway and lipid biosynthesis
Mechanistic study of human TPI deficiency
Features & Benefits • Simple, Sensitive & Rapid protocol
• Highly-Sensitive: Detects as low as 0.5 µM DHAP
Kit Components • DHAP Assay Buffer
• PicoProbe™ (in DMSO)
• DHAP Enzyme Mix
• DHAP Developer
• DHAP Standard
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Dihydroxyacetone Phosphate (DHAP) is an important intermediate in both lipid biosynthesis and glycolysis. In glycolysis, Fructose-1,6-diphosphate is converted to DHAP and glyceraldehyde-3-phosphate (GAP) by aldolase. Both DHAP and GAP serve as the intracellular triose phosphate pool. DHAP can be further converted into GAP by Triose Phosphate Isomerase (TPI). In humans, TPI deficiency is a rare autosomal disease. It causes hemolytic anemia, neurological diseases, and even death due to blockage of the glycolytic pathway and accumulation of DHAP in erythrocytes. Biovision’s PicoProbe™ Dihydroxyacetone Phosphate Assay kit is suitable for measuring low levels of DHAP typically found in a variety of samples. In this kit, TPI converts DHAP to GAP that undergoes a series of reaction and reduces PicoProbe™ to generate fluorescence. The fluorescence intensity generated is directly proportional to the amount of Dihydroxyacetone Phosphate. This simple, sensitive, and easy to use assay kit can detect Dihydroxyacetone Phosphate as low as 0.5 µM in a variety of samples.


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Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
BioVision kits are optimized 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 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. But once the samples are deproteinized, this is not applicable.
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.