Renin, a highly specific aspartyl protease, cleaves angiotensinogen to yield angiotensin I, which is further converted into angiotensin II by ACE (Angiotensin Converting Enzyme). Angiotensin II constricts blood vessels leading to increased blood pressure. It also increases the secretion of ADH and aldosterone, and stimulates the hypothalamus to activate the thirst reflex. Since an overactive renin-angiotensin system leads to hypertension, renin is an attractive target for the treatment of this disease. The SensoLyte® 390 Renin Assay Kit provides a convenient assay for high throughput screening of renin inhibitors and for continuous assay of renin activity using a Mc-Ala/Dnp fluorescence resonance energy transfer (FRET) peptide. In the FRET peptide the fluorescence of Mc-Ala is quenched by Dnp. Upon cleavage into two separate fragments by renin, the fluorescence of Mc-Ala is recovered, and can be monitored at excitation/emission = 330/390 nm. This assay is about four fold more sensitive than an EDANS/DABCYL-based assay and can detect 8 ng/ml of renin.
Component A: Renin substrate Mc-Ala/Dnp FRET peptide Ex/Em=330 nm/390 nm upon cleavage: 2 mM DMSO solution, 50 µL
Component B: Mc-Ala fluorescence reference standard Ex/Em=330 nm/390 nm: 3 mM, 5 µL
Component C: Human recombinant renin: 100 µL
Component D: Assay buffer: 25 mL
Component E: Renin Inhibitor Ac-HPFV- (Sta)-LF-NH2: 1mM DMSO solution, 5 µL
Storage & stability
Store all kit components at -20°C, except for Component C. Store Component C at -80°C. Aliquots are needed to avoid multiple freeze-thaw cycles. Protect Components A and B from light and moisture. Component D can be stored at room temperature for convenience.