Recombinant Mouse Prostasin/Prss8 Protein, CF

R&D Systems | Catalog # 2968-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Prostasin/Prss8 Protein (2968-SE)
  • 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

Q99L44

Applications

Enzyme Activity
View all Prostasin/Prss8 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Prostasin/Prss8 protein
Ala30-Gln289 with an N-terminal signal sequence and 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

Ala30 & Ile45

Predicted Molecular Mass

29 kDa & 28 kDa

SDS-PAGE

37-39 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate Boc-QAR-AMC (Catalog # ES014).
The specific activity is >15 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

2968-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2.
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: Prostasin/Prss8

Prostasin encoded by the Prss8 gene, also known as channel activating protease 1, is a serine protease with a trypsin-like substrate specificity (1, 2). The preproenzyme possesses a C-terminal membrane-spanning domain that can be proteolytically processed to generate a secreted form of the enzyme. The secreted form of prostasin has been purified from seminal fluid (2). Prostasin is highly expressed in the prostate gland, and is expressed at lower levels in the lung, kidney, liver, salivary gland, and pancreas (3). The peptidase activity of prostasin is involved in the regulation of epithelial sodium channels (4, 5). Secreted from NS0 cells as the active form, the purified recombinant mouse (rm) Prostasin most likely consists of a disulfide bond-linked two chains with N-terminal sequences of I45TGGGSAKPG and A30DGTE, respectively (3). The enzymatic activity of rmProstasin is effectively inhibited by rmHAI-1 and rmHAI‑2B (Catalog # http://www.rndsystems.com/product_results.aspx?k=1141-PI">1141-PI and http://www.rndsystems.com/product_results.aspx?k=1107-PI">1107-PI).

References

  1. Vuagniaux, G. et al. (2000) J. Am. Soc. Nephrol. 11:828.
  2. Yu J.X. et al. (1994) J. Biol. Chem. 269:18843.
  3. Yu J.X. et al. (1995) J. Biol. Chem. 270:13483.
  4. Caldwell R.A. et al. (2004) Am. J. Physiol. Cell Physiol. 286:C190.
  5. Tong Z. et al. (2004) Am. J. Physiol. Lung Cell Mol. Physiol. 287:L928.

Alternate Names

Prss8

Entrez Gene IDs

5652 (Human); 76560 (Mouse)

Gene Symbol

PRSS8

UniProt

Q99L44

Additional Prostasin/Prss8 Products

Product Documents for Recombinant Mouse Prostasin/Prss8 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 Prostasin/Prss8 Protein, CF

For research use only

Citations for Recombinant Mouse Prostasin/Prss8 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Prostasin/Prss8 Protein, CF (2968-SE):

Materials
  • Assay Buffer: 50 mM Tris, 0.05% (w/v) Brij-35, pH 9.0
  • Recombinant Mouse Prostasin/Prss8 (rmProstasin) (Catalog # 2968-SE)
  • Substrate: BOC-Gln-Ala-Arg-AMC (Catalog # ES014)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmProstasin to 20 µg/mL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Load into a black well plate 50 µL of 20 µg/mL rmProstasin and start the reaction by adding 50 µL of 200 µM Substrate. As a Substrate Blank combine 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmoles/min/µg) =

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

     *Adjusted for Substrate Blank
     **Derived using calibration standard 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

Per Well:

  • rmProstasin: 1 µg
  • Substrate: 100 µM

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