Isocitrate Assay Kit
ISOCITRATE (ISOCITRIC ACID) is a substrate in the citric acid (TCA) cycle. Isocitrate is formed by the isomerization of citrate catalyzed by the enzyme aconitase. Isocitrate is oxidized by isocitrate dehydrogenase producing alpha-ketoglutarate and generating NADPH. Isocitrate is commonly found in many fruits and vegetables and their processed products. Industrially, isocitrate is used as a marker to identify the quality and purity of fruit juices.Assay Genie's isocitrate assay measures the NADPH generated from the oxidation of isocitrate. The NADPH reduces a probe into a highly fluorescent product. The fluorescence intensity of this product, measured at Ex/Em = 530/585 nm, is proportional to the isocitrate concentration in the sample.
ApplicationsFor quantitative determination of isocitrate and evaluation of drug effects on its metabolism.
- Fast and sensitive. Linear detection range (20 uL sample): 0.6 to 500 uM for 10 min reaction.
- Convenient and high-throughput. Homogeneous "mix-incubate-measure" type assay. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.
Isocitrate Assay Kit Information:
|Kit Includes:||Assay Buffer: 10 mL Enzyme A: 120 uL NADP: 500 uL Enzyme B: 120 uL Standard: 1 mL Probe: 750 uL|
|Kit Requires:||Pipetting devices and accessories (e.g. multi-channel pipettor), black flat-bottom 96-well plates, centrifuge tubes and fluorescence plate reader capable of reading at EX/EM= 530/585 nm.|
|Method of Detection:||FL530/585 nm|
|Detection Limit:||0.6 uM|
|Samples:||Food, beverage, biological samples (e.g. cell lysate, tissue homogenate, serum, etc.)|
|Protocol Length:||10 min|
|Storage:||Store all components at -20°C upon receiving|
|Shelf Life:||6 months|