Recombinant Human Prostasin/Prss8 Protein, CF Summary
Product Specifications
Ala33-Gly319, with a C-terminal 10-His tag
The pro form was activated and further purified.
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.
4599-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 9.5
- Recombinant Human Prostasin/Prss8 (rhProstasin) (Catalog # 4599-SE)
- Substrate: BOC-Gln-Ala-Arg-AMC (Catalog # ES014), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhProstasin to 20 µg/mL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load 50 µL of 20 µg/mL of rhProstasin into a well, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 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 (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard 7-amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891).
Per Well:- rhProstasin: 1 µg
- Substrate: 100 µM
Reconstitution Calculator
Background: Prostasin/Prss8
Prostasin, also known as channel activating protease 1, is a serine protease with 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).
- 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:C194.
- Tong, Z. et al. (2004) Am. J. Physiol. Lung Cell Mol. Physiol. 287:L928.
Citations for Recombinant Human 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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Citations: Showing 1 - 2
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Reduced prostasin (CAP1/PRSS8) activity eliminates HAI-1 and HAI-2 deficiency-associated developmental defects by preventing matriptase activation.
Authors: Szabo, Roman, Uzzun Sales, Katiuchi, Kosa, Peter, Shylo, Natalia, Godiksen, Sine, Hansen, Karina K, Friis, Stine, Gutkind, J Silvio, Vogel, Lotte K, Hummler, Edith, Camerer, Eric, Bugge, Thomas H
PLoS Genet, 2012-08-30;8(8):e1002937.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Matriptase initiates activation of epidermal pro-kallikrein and disease onset in a mouse model of Netherton syndrome.
Authors: Sales KU, Masedunskas A, Bey AL
Nat. Genet., 2010-07-25;42(8):676-83.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay
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