Recombinant Mouse Prostasin/Prss8 Protein, CF Summary
Product Specifications
Ala30-Gln289 with an N-terminal signal sequence and a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
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.
|
Assay Procedure
- 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
- Dilute rmProstasin to 20 µg/mL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- 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.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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).
- rmProstasin: 1 µg
- Substrate: 100 µM
Reconstitution Calculator
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 # 1141-PI and 1107-PI).
- Vuagniaux, G. et al. (2000) J. Am. Soc. Nephrol. 11:828.
- Yu J.X. et al. (1994) J. Biol. Chem. 269:18843.
- Yu J.X. et al. (1995) J. Biol. Chem. 270:13483.
- Caldwell R.A. et al. (2004) Am. J. Physiol. Cell Physiol. 286:C190.
- Tong Z. et al. (2004) Am. J. Physiol. Lung Cell Mol. Physiol. 287:L928.
Citations for Recombinant Mouse Prostasin/Prss8 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Mutational tail loss is an evolutionary mechanism for liberating marapsins and other type I serine proteases from transmembrane anchors.
Authors: Raman K, Trivedi N, Raymond W, Ganesan R, Kirchhofer D, Verghese G, Craik C, Schneider E, Nimishakavi S, Caughey G
J Biol Chem, 2013-02-27;288(15):10588-98.
Species: Mouse
Sample Types: Recombinant Protein
Applications: Enzyme Assay -
Structure of human prostasin, a target for the regulation of hypertension.
Authors: Rickert KW, Kelley P, Byrne NJ, Diehl RE, Hall DL, Montalvo AM, Reid JC, Shipman JM, Thomas BW, Munshi SK, Darke PL, Su HP
J. Biol. Chem., 2008-10-14;283(50):34864-72.
Species: Human
Sample Types:
Applications: Enzyme Assay
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