Human Renin Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human Renin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human mature Renin and Prorenin in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human Renin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Renin showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Renin.

Preparation and Storage

Background: Renin

Renin is a secreted endopeptidase that plays a crucial role in the regulation of blood pressure and salt balance through the cleavage of angiotensinogen, which is the only known physiological substrate of Renin. The 406 aa precursor includes a signal peptide (aa 1 - 23), a propeptide (aa 24 - 66), and a mature chain (aa 67 - 406). The amino acid sequence of human Renin is 100%, 73%, 71% and 67% identical to that of chimpanzee, canine, mouse and rat.

Mouse Renin is a secreted, 42 - 47 kDa glycosylated member of the peptidase A1 family. It is an aspartyl protease that cleaves angiotensinogen to form angiotensin I. In mouse, there are two genes that code for Renin. One is in the submandibular gland and the other is in the kidney. The two mature Renin molecules are 95% amino acid (aa) identical. Renal Renin (Renin-1) is synthesized as a 381 aa proform (aa 22 - 402). In the kidney, prorenin is proteolytically cleaved after Thr71 to generate a mature enzyme. Mouse pro-renin shares 70% and 85% aa sequence identity with human and rat pro-renin, respectively.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1、Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2、Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3、Add 50 μL of Assay Diluent to each well.

4、Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5、Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6、Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7、Aspirate and wash 5 times.

8、Add 100 μL Substrate Solution to each well.

9、Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.