Angiotensin Converting Enzyme 2 Activity, Fluorimetric 390, BioAssay Kit (ACE-2, ACEH)

Angiotensin I converting enzyme 2 (ACE2), the newest member of the renin angiotensin system (RAS), is a zinc metallopeptidase that plays a central role in the control of angiotensin peptides (1,2). ACE2 has direct effects on cardiac function (3), and is expressed predominantly in vascular endothelial cells of the heart and the kidneys (2). It has been reported that ACE2 might protect kidneys in early stages of diabetes (4). In addition to its role in the regulation of hypertension, ACE2 is a functional receptor for coronavirus that causes severe acute respiratory syndrome (SARS) (5). ACE2 is considered an important therapeutic target for controlling cardiovascular diseases, kidney disease and severe acute respiratory syndrome (SARS) outbreaks., The Fluorometric ACE2 Activity Assay Kit provides a convenient assay for high throughput screening of ACE2 inhibitors and inducers and for continuous assay of ACE2 activity using Mca/Dnp fluorescence resonance energy transfer (FRET) peptide. In the FRET peptide the fluorescence of Mca is quenched by Dnp. Upon cleavage into two separate fragments by the enzyme, the fluorescence of Mca is recovered, and can be monitored at excitation/emission = 330/390nm. The assay can detect the activity of subnanogram level of ACE2. Assays are performed in a convenient 96-well microplate format. Kit Components: A2295-04B1: Mca/Dnp, ACE2 Substrate, Ex/Em=330nm/390nm upon cleavage 5mM, 1x50ul, A2295-04B2: Mca Fluorescence Reference Standard, Ex/Em=330nm/390nm, 1mM, 1x10ul, A2295-04B3: Assay Buffer 1x20ml, A2295-04B4: Inhibitor of ACE2, 100uM, 1x10ul, A2295-04B5: Stop Solution, 1x10ml, Storage and Stability: Store all kit components at -20°C. Unused kit is stable for 6 months after receipt. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.  Protect A2295-04B1 and A2295-04B2 from light and from moisture.  A2295-04B3 and A2295-04B5 can be stored at room temperature for convenience. Materials Required But Not Provided:  ACE2 source: The active enzyme, cell lysates, tissue extracts.,  96-well microplate: Black microplates provide better signal to noise ratio.,  Fluorescence microplate reader: Capable of detecting emission at 390nm with excitation at 330nm. Quality Control: As reported