Non radioactive HPRT assayPRECICE® HPRT Assay kit provides the first non radioactive protocol for measurement of HPRT activity in a convenient 96-well plate format.Hypoxanthine-guanine phosphoribosyltransferase (HPRT, EC 2.4.2.8), a key enzyme of the purine salvage pathway, is encoded by highly variable HPRT1 gene. More than 300 mutations in HPRT1 gene associated with genetic disorders in humans have been described leading to partial or complete deficiency of HPRT enzyme. Complete HPRT deficiency is a rare human genetic disorder, also known as Lesch-Nyhan syndrom. In some cases, gout, more frequent purine disorder, can also be secondary to HPRT deficiency (Ceballos-Picot I. et al 2010). High variability of HPRT1 gene complicates genetic testing. Currently, HPRT enzyme functions are tested in red blood cell lysates in a chromatographic assay using radiolabeled 14C-hypoxanthine (Cartier P. et al, 1968). Principle Wide linearity range
Accurate
Fast
Convenient
Validated
|
V mouse over to enlarge
|
Calibration curve of IMPDH-based PRECICE® HPRT Assay
The rate of the increase in absorbance at 340nm per hour as a function of the concentration of human recombinant HPRT enzyme (NovoCIB, ref. E-Nov-9). The changes in absorbance corresponding to 3 different control hemolysates diluted in complete reaction mixture to final hemoglobin concentration 1mg/ml (n=4) are shown by filled squares, filled triangles and filled circles. The insert shows a linear correlation observed in whole range of 21ng/ml up to 1.5�g/ml of recombinant HPRT; the units are the same. |
References |
Related Links | |
• HGPRT enzyme • Bacterial Recombinant IMPDH |