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Renin (human), (recombinant) (active)

Highly specific active enzyme cleaves angiotensinogen to yield angiotensin I
 
ENZ-PRT193-0010 10 µg 112.00 USD
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Product Specification

Alternative Name:Angiotensinogenase, Angiotensin forming enzyme, Ren
 
MW:~52kDa (predicted is 38.3 kDa)
 
Source:Produced in HEK cells. Mature Renin (aa 67-406) is fused at the N-terminus to a FLAG®-tag. Active form, does not contain prorenin peptide (aa 1-66).
 
UniProt ID:P00797
 
Concentration:0.1 mg/ml when reconstituted as suggested.
 
Formulation:Lyophilized. In 25mM MES buffer containing 150mM NaCl.
 
Purity:≥90%
 
Purity Detail:Affinity purifed.
 
Endotoxin Content:<0.1EU/µg protein (LAL test)
 
Specific Activity:≥30U/mg. 1 unit is defined as the amount of enzyme that cleaves 1 pmole of the fluorogenic peptide substrate, Arg-Glu(EDANS)-Ile-His-Pro-Phe-His-Pro-Phe-His-Leu-Val-Ile-His-Thr-Lys(dabcyl)-Arg.
 
Shipping:Shipped on Blue Ice
 
Short Term Storage:-20°C
 
Long Term Storage:-20°C
 
Use/Stability:Reconstitute in 100µl deionized H2O for a final concentration of 0.1mg/ml.
 
Handling:Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and store at -80°C.
 
Scientific Background:Renin is a highly specific aspartyl protease that participates in the body's renin-angiotensin system (RAS). It cleaves angiotensinogen, which is produced in the liver, to yield angiotensin I. This is further converted into angiotensin II by angiotensin converting enzyme. Angiotensin II has been shown to constrict blood vessels and increase sodium reabsorption in the kidneys, leading to increased blood pressure. The primary structure of renin precursor consists of 406 amino acids with a pre- and a pro-segment carrying 20 and 46 amino acids, respectively. Mature renin contains 340 amino acids and has a predicted mass of ~37 kDa.
 
Technical Info/Product Notes:FLAG® is a registered trademark of Sigma-Aldrich Co.
 
ENZ-PRT193 SDSPAGE
Figure 1. Coomassie stained SDS-PAGE. Lane 1, Molecular weight marker. Lane 2, 1.0µg Human active renin (ENZ-PRT193).
ENZ-PRT193 Activity Assay
Figure 2. Renin activity using the FRET peptide substrate, Arg-Glu(EDANS)-Ile-His-Pro-Phe-His-Pro-Phe-His-Leu-Val-Ile-His-Thr-Lys (DARBCYL)-Arg. Briefly, a 10µg vial of renin (ENZ-PRT193) was resuspended in 100µl dH2O. The renin protein was diluted to 20µg/ml in 50µl of buffer (25mM MES pH6 and 150mM NaCl). Separately, the renin substrate was diluted to 20µM in a 50µl volume. Both the enzyme and substrate were heated to 37°C for 10 minutes. They were then combined and read kinetically every minute for 10 minutes (using Ex350nm / Em 490nm). Interpolation off a standard curve (calibration standard Fmoc-Glu(EDANS)-OH) was used to calculate the activity of the renin enzyme. The activity was calculated to be > 30pmole/µg/min.
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ENZ-PRT193 SDSPAGE ENZ-PRT193 Activity Assay

General Literature References

Catalytic Properties of Intramembrane Aspartyl Protease Substrate Hydrolysis Evaluated Using a FRET Peptide Cleavage Assay: S.H. Naing; ACS Chem. Biol. 10, 2166 (2015),
Cloning and sequence analysis of cDNA for human renin precursor: T. Imai, et al.; PNAS 80, 7405 (1983), Abstract;
Renin-angiotensin system: biochemistry and mechanisms of action: M.J. Peach; Physiol. Rev. 57, 313 (1977), Abstract;

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