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Glycerol-3-Phosphate (G3P) Colorimetric Assay Kit

Catalog #: K641

In stock

$420.00

Product Details

Cat # +Size K641-100
Size 100 assays
Detection Method Absorbance (450 nm)
Applications Measurement of Glycerol-3-Phosphate in various tissues/cells
Analysis of carbohydrate and lipid metabolism and cell signaling
Drug screening
Features & Benefits • Simple, sensitive, and rapid.
• This assay kit can detect G3P less than 2 nmol/well and can be used for a variety of sample types.
Kit Components • G3P Assy Buffer
• G3P Enzyme Mix (Lyophilized)
• G3P Probe (Lyophilized)
• G3P Standard (Lyophilized)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Glycerol-3-phosphate (G3P) is an important intermediate for all living organisms. Glycerol-3-Phosphate is produced either by glycerol via glycerol kinase or by dihydroxyacetone phosphate through glycerol-3-phosphate dehydrogenase. In response to cellular signals, glycerol-3-phosphate can be utilized in multiple pathways: it can be further converted into glyceraldehyde-3-phosphate and enter glycolysis or rapidly generate NAD+ in brain or muscle tissues through the G3P shuttle or enter the lipid biosynthetic pathway. Recent studies have found that glycerol-3-phosphate is a novel regulator and plays a fundamental defense role in plant pathogenesis. BioVision’s Glycerol-3-Phosphate Assay kit is a sensitive, fast and easy-to-use kit. In this assay, G3P is oxidized by G3P Enzyme Mix to form an intermediate, which reduces a nearly colorless probe to a colored product with strong absorbance at 450 nm. This assay kit can detect G3P less than 2 nmol/well and can be used for a variety of sample types.


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There is high background, close to OD=0.5. What can be done to remove this?
NADH in samples can create background. The background values need to subtracted from the G3P reaction values. Sometimes, diluting the sample helps since the background gets diluted while the analyte is still within the range of the standard curve. This is a delicate balance where the best results are obtained.
Can heparin be used for blood collection for this assay?
Yes, heparin should be fine.
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 as long as there is active G3PDH in the sample. 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.
Does the slope of the standard curve in all experiments need to be the same? What is the acceptable variability?
The R squared value signifies the probability that the data points are on a straight line. As long as this value sis close to 98 or 99%, the std curve can be trusted. The slopes can vary depending on the instrument, the dilutions used etc.
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 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.
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