Recombinant Mouse Renin Protein, CF

R&D Systems | Catalog # 4277-AS

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Renin Protein (4277-AS)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

P06281

Applications

Enzyme Activity
View all Renin Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Renin protein
Leu22-Arg402, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Leu22

Predicted Molecular Mass

43 kDa

SDS-PAGE

45-55 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate 5-FAM/QXL™ 520. Peti-Peterdi, J. et al. (2009) Physiology 24:88.
The specific activity is >20 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

4277-AS
Formulation Supplied as a 0.2 μm filtered solution in MES and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Renin

Renin is a aspartyl protease that plays a crucial role in the regulation of blood pressure and salt balance through the renin-angiotensin system (1). It cleaves angiotensinogen to generate angiotensin I, which can be further converted by angiotensin converting enzyme (ACE) to angiotensin II. Angiotensin II is the active molecule of the renin-angiotensin system that acts by binding to angiotensin receptors type 1 and 2 (AT-1 and AT-2), and has direct pathophysiological effects on the heart and peripheral vasculature (2). After secretion, inactive prorenin can be proteolytically activated by trypsin, cathepsin B, or other proteinases. Because of its unique specificity, inhibition of Renin is widely studied to provide selective therapy for hypertension, congestive heart failure and associated disorders (3).

References

  1. Fuminaki, S. et al. 2004 in Handbook of Proteolytic Enzymes, Barrett, A. J. et al. eds:54.
  2. Unger, T. et al. 2008 Medscape J. Med. 10:S4.
  3. Dhanaraj, V. et al. 1992 Nature 357:466.

Alternate Names

Angiotensinogenase, REN

Entrez Gene IDs

5972 (Human); 19701 (Mouse)

Gene Symbol

REN

UniProt

P06281

Additional Renin Products

Product Documents for Recombinant Mouse Renin Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse Renin Protein, CF

QXL is a trademark of AnaSpec, Inc.

For research use only

Citations for Recombinant Mouse Renin Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Renin Protein, CF (4277-AS):

Materials
  • Activation Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl2, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Assay Buffer: 50 mM NaH2PO4, 1 M NaCl, pH 7.5
  • Recombinant Mouse Renin (rmRenin) (Catalog # 4277-AS)
  • Trypsin (Sigma, Catalog # T-1426)
  • AEBSF (Catalog # EI001), 100 mM stock in DMSO
  • Substrate: “Rat Renin FRET” 5-FAM/QXL™520 (AnaSpec, Catalog # 62334), 100 µM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Activate rmRenin at 0.1 mg/mL with Trypsin at 2 µg/mL.
    1. Dilute rmRenin to 0.2 mg/mL in Activation Buffer.
    2. Dilute Trypsin to 4 µg/mL in Activation Buffer.
    3. Mix equal volumes of the diluted rmRenin and Trypsin.
    4. Incubate at 37 °C for 1 hour.
  2. Stop Trypsin activity with 1 mM AEBSF.
    1. Dilute AEBSF to 2 mM in Assay Buffer.
    2. Add a volume of 2 mM AEBSF equal to the reaction volume for 1 mM AEBSF and 50 µg/mL rmRenin.
    3. Incubate at room temperature for 30 minutes.
  3. Dilute Substrate to 4 µM in Assay Buffer.
  4. Dilute rmRenin to 2 ng/µL in Assay Buffer.
  5. Load into a black well plate 50 µL of 2 ng/µL rmRenin (100 ng/well) and start the reaction by adding 50 µL of 4 µM Substrate.
  6. Read at excitation and emission wavelengths of 490 nm and 520 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard calibration standard 5-FAM (AnaSpec, Catalog # 60584).

  • rmRenin: 0.1 µg
  • Substrate: 2 µM

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