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Isocitrate Colorimetric Assay Kit

Catalog #: K656
$370.00

Product Details

Cat # +Size K656-100
Size 100 assays
Detection Method Absorbance (450 nm)
Species Reactivity Mammalian
Applications Isocitrate Assay Kit can detect 1 to 20 nmoles (~0.2 – 5 µg) of isocitrate.
Features & Benefits • Simple procedure; takes ~ 40 minutes
• Fast and convenient
• Kit contains all necessary reagents for accurate measurement of isocitrate levels
Kit Components • Isocitrate Assay Buffer
• Isocitrate Enzyme Mix
• Substrate Mix
• Isocitrate Standard (100 mM)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Isocitric acid (HOOC-CHOH-CH (-COOH)-CH₂-COOH) is an intermediate of the Krebs TCA cycle, positioned between citrate and α-keto-glutarate. It is the branch point from which the glyoxylate shunt operates in plants and lower organisms. Isocitrate is found in substantial concentrations in many fruits and vegetables as well as in foods produced from these raw materials. In the TCA cycle, isocitrate is oxidized by isocitrate dehydrogenase (IDH) to α-ketoglutarate with the generation of NAD(P)H. Loss of NAD-IDH has been implicated as a potential causative factor in retinitis pigmentosa. BioVision's Isocitrate Assay Kit provides a simple, sensitive and rapid means of quantifying isocitrate in a variety of samples. In the assay, isocitrate is oxidized with the generation of NADPH which converts a nearly colorless probe to an intensely colored species with a λmax of 450nm. The Isocitrate Assay Kit can detect 1 to 20 nmoles (~0.2 – 5 µg) of isocitrate.


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What does the enzyme mix in the kit do?
The enzyme mix contains enzymes that catalyze the detection reactions leading to measurement of Isocitrate in the sample. Without the enzymemix the background NADH and NADPH can be detected.
Does this kit also detect citrate?
No, the enzyme at the first step of the detection process is very specific and only reacts with isocitrate.
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. 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.
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.