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Product Details
| Cat # +Size | K420-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Fluorescence (Ex/Em 535/587nm) |
| Species Reactivity | Mammalian |
| Applications | Measure Phosphate concentrations between 2 µM and 10 µM, with a lower detection limit of approximately 100 pmol. |
| Features & Benefits | • Simple procedure; takes ~ 3 hours • Fast and convenient • The assay is sensitive and stable • The TUNEL-based assay kit provides complete components including positive and negative control cells for convenient detection of DNA fragmentation in cultured cells and tissue sections. |
| Kit Components | Phosphate Assay Buffer PicoProbe™ (in DMSO) Converter Developer Phosphate Substrate |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Inorganic phosphate (Pi) is one of the most important ions in biological systems. It functions in a variety of roles. One of its most important roles is as a molecular switch, turning enzyme activity on and off through the mediation of the various protein kinases and phosphatases in biological systems. A highly sensitive assay is desired to monitor Pi in a variety of samples. The newly designed and enhanced Phosphate Assay Kit provides a sensitive, easy and quick means of assessing Pi over a wide range of concentrations. In this assay, inorganic phosphate reacts with sucrose to produce glucose-1-phosphate in the presence of a proprietary enzyme. The glucose-1-phosphate is specifically oxidized to generate a product that reacts with the PicoProbe™ to generate fluorescence (Ex/Em = 535/587 nm). The kit can be used to detect Pi produced through reactions involving ATPases, GTPases, 5’-nucleotidase, protein phosphatases, acid and alkaline phosphatases, phosphorylase, etc. from a variety of samples. Unlike other commercially available assays, this assay is not affected by the presence of glucose in samples. This kit can determine phosphate concentrations between 2 µM and 10 µM, with a lower detection limit of approximately 100 pmol.
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Is it possible to quantify the phosphate in an acidic solution (Meaning 1M HCl) using this kit?
The assay is designed to function in neutral pH. The enzymes in the Maltose converter and glucose developer will likely not work at extreme acidic pH. Also the probe will likely not work. We have not tested this assay with 1M HCl.
What would you consider to be a significant amount of glucose? In our red blood cell samples, for example, there might be up to 30 mmol/L glucose. Would this be enough to warrant a correction?
if there is glucose in the samples in comparable amounts with the phosphate (no specific amount can be suggested, depends upon relative amounts of glucose and phosphate in the sample), a glucose control is recommended for the purpose of proper background subtraction. The fluorometric kit being 10-fold more sensitive, the background can be amplified many times.
What is the difference between K410-100 and K420-100?
K410 is a colorimetric assay with a working range of 1-10uM concentration of phosphate. K410-550 provides enough materials to carry out 500 assays.K420 is a more sensitive fluorometric assay with a working range of 0.4-10uM. K420-100 provides materials to perform 100 assays.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only.
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 your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
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.
Erkizan et al., RNA helicase A activity is inhibited by oncogenic transcription factor EWS-FLI1. Nucleic Acids Res., Jan 2015; 43: 1069 - 1080.
For more citations of this product click here
