Renin Inhibitor Screening Assay Kit

Cayman Chemical Item Number 10006270

Renin Inhibitor Screening Assay Kit

Description

Renin is an aspartyl protease of approximately 40 kDa that is released in active form from renal juxtaglomerular cells in response to sodium depletion, decreased blood volume and blood pressure, and b-adrenergic stimulation.^1,2,3 Renin catalyzes the initial and rate limiting step in the renin-angiotensin system (RAS) pathway, converting angiotensinogen into angiotensin I. Angiotensin Converting Enzyme (ACE) subsequently converts angiotensin I to angiotensin II, which is a potent vasoconstrictor. The Cayman Chemical Renin Inhibitor Screening Assay Kit provides a convenient assay in a 96-well format for evaluating human renin inhibitors. The assay utilizes a renin-based synthetic peptide substrate which incorporates the fluorophore EDANS at one end and an EDAN-quenching molecule (Dabcyl) at the other end.⁴ After cleavage by renin, the peptide-EDANS product is released yielding bright fluorescence which can be easily analyzed using excitation wavelengths of 335-345 nm and emission wavelengths of 485-510 nm. The assay kit includes recombinant human renin (sufficient for 100 reactions), substrate, buffers, and complete instructions.

1 Imai, T., Miyazaki, H., Hirose, S., et al. Cloning and sequence analysis of cDNA for human renin precursor. Proc Natl Acad Sci USA 80 7405-7409 (1983).

2 Cartledge, S., and Lawson, N. Aldosterone and renin measurements. Ann Clin Biochem 37 262-278 (2000).

3 Persson, P.B. Renin: Origin, secretion and synthesis. J Physiol 552.3 667-671 (2003).

4 Wang, G.T., Chung, C.C., Holzman, T.F., et al. A continuous fluorescence assay of renin activity. Anal Biochem 210 351-359 (1993).

Stability

1 year

Storage

-80°C

Shipping

Dry ice in continental US; may vary elsewhere

Background Reading

Imai, T., Miyazaki, H., Hirose, S., et al. Cloning and sequence analysis of cDNA for human renin precursor. Proc Natl Acad Sci USA 80 7405-7409 (1983).

Persson, P.B. Renin: Origin, secretion and synthesis. J Physiol 552.3 667-671 (2003).

Wang, G.T., Chung, C.C., Holzman, T.F., et al. A continuous fluorescence assay of renin activity. Anal Biochem 210 351-359 (1993).

Wood, J.M., Gulati, N., Forgiarini, P., et al. Effects of a specific and long-acting renin inhibitor in the marmoset. Hypertension 7 797-803 (1985).

Holzman, T.F., Chung, C.C., Edalji, R., et al. Recombinant human prorenin from CHO cells: Expression and purification. J Protein Chem 9(6) 663-672 (1990).

Cartledge, S., and Lawson, N. Aldosterone and renin measurements. Ann Clin Biochem 37 262-278 (2000).

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10006270-96WELL

Renin Assay Buffer (10X)

1 ea

Renin (human recombinant) Assay Reagent

1 ea

Renin Substrate

1 ea

96-Well Solid Plate (black)