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PicoProbe™ Fructose-6-Phosphate Fluorometric Assay Kit

based on 1 citations in multiple journalsPicoProbe™ Fructose-6-Phosphate Fluorometric Assay Kit14.1 4
Highly Sensitive Assay
Catalog #: K689
$460.00

Product Details

Cat # +Size K689-100
Size 100 assays
Detection Method Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The PicoProbe™ F6P Assay Kit can detect F6P in the range of 0.01 to 0.5 nmoles with detection sensitivity ~1 µM of F6P.
Features & Benefits • Simple procedure; takes ~ 35-40 minutes to complete the assay
• Fast and convenient
• Sensitive, efficient and accurate assay
Kit Components • F6P Assay Buffer
• F6P Probe
• F6P Enzyme Mix
• F6P Developer
• F6PStandard (10 µmol)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Fructose-6-phosphate (F6P) is an important intermediate in the glycolytic pathway which leads from glucose to pyruvate. It is formed from glucose-6-phosphate and is further phosphorylated to fructose-1,6-diphosphate which is subsequently cleaved to glyceraldehyde phosphate and dihydroxyacetone phosphate. The transformation of glucose-6-phosphate is controlled by phosphoglucose isomerase, a very interesting enzyme in that it possesses multiple functions, as an isomerase, a neuroleukin, an autocrine motility factor and a differentiation and maturation mediator (1). BioVision's PicoProbe ™ Fructose-6-phosphate Assay Kit is a fluorescence-based simple, highly sensitive and rapid means of quantifying F6P in a variety of samples. In the assay, F6P is converted to glucose-6-phosphate which is subsequently oxidized with the generation of a fluorescent product. The PicoProbe™ F6P Assay Kit can detect F6P in the range of 0.01 to 0.5 nmoles with detection sensitivity ~1 µM of F6P.


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There is no difference between the deproteinized sample and its corresponding background control. What could be the reason?
Our thinking is that there is much more NADH, NADPH and G6P combined in the samples than F6P. Also, the F6P might be getting broken down by cellular enzymes. So it is important to keep the cells on ice the whole time. We sugegst the following:
-Pellet cells and remove all liquid. Add a small volume 20-50ul of water and resuspend.
- Add PCA immediately and continue the deproteinization. Collect the supernatant after the PCA precipitation and neutralization. Use this for the subsequent assay.
- This way the samples will be more concentrated. I would suggest trying several different volumes to see which one yields the best readings.
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.
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.
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