Intended UseHuman REN precursor ELISA Kit allows for the in vitro quantitative determination of REN precursor , concentrations in serum, Plasma , tissue homogenates and Cell culture supernates and Other biological fluids.
StorageFor 5-7days:Store the whole kit at 4℃
For a Long time :Store the Substrate at 4℃, other reagent should store at -20℃.
Product Description specificalIntroduction: The biosynthesis of renin in the mouse submaxillary gland was defined using both cell-free translation and pulse-labelling methods. Renin is synthesized as a preproform and cleaved by microsomes to the proform. Prorenin (MW = 46 kilodaltons, pI 6.35) is processed intracellulary into an intermediate form (MW = 41 kilodaltons). The latter is converted by a slow intracellular process to the final major storage form (MW = 37 kilodaltons). Both the intermediate and the 37 KD forms are secreted. The biosynthetic precursor does not bind to pepstatin-aminohexyl sepharose. In parallel experiments, using rapid tissue extraction with buffers containing protease inhibitors and fractionation with affinity chromatography, we have also identified in the mouse submaxillary gland an inactive form of renin which has a MW of 48 2 KD and pI of 6.4. Mouse submaxillary gland inactive renin binds to affigel-blue and not to pepstatin-sepharose. The similarity in the properties of inactive renin and the biosynthetic renin precursor lends further support to the suggestion that tissue inactive renin may be the renin precursor
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to renin precursor. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for renin precursor and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain renin precursor, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of renin precursor in the samples is then determined by comparing the O.D. of the samples to the standard curve.