Recombinant Human Prostasin/Prss8 Protein, CF

Catalog # Availability Size / Price Qty
4599-SE-010
R&D Systems Recombinant Proteins and Enzymes
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Citations (2)
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Recombinant Human Prostasin/Prss8 Protein, CF Summary

Product Specifications

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.
Activity
Measured by its ability to cleave the fluorogenic peptide substrate Boc-QAR-AMC (Catalog # ES014). The specific activity is >10 pmol/min/µg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived human Prostasin/Prss8 protein
Ala33-Gly319, with a C-terminal 10-His tag
The pro form was activated and further purified.
Accession #
N-terminal Sequence
Analysis
Ala33 & Ile45
Structure / Form
Active
Predicted Molecular Mass
32 kDa (Pro) & 31 kDa (Mature)
SDS-PAGE
39 kDa, reducing conditions

Product Datasheets

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4599-SE

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • 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
  1. Dilute rhProstasin to 20 µg/mL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. 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.
  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 (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

Reconstitution Calculator

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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).

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:C194.
  5. Tong, Z. et al. (2004) Am. J. Physiol. Lung Cell Mol. Physiol. 287:L928.
Entrez Gene IDs
5652 (Human); 76560 (Mouse)
Alternate Names
CAP1; channel-activating protease 1; EC 3.4.21; EC 3.4.21.-; EC 3.4.21.120; Prostasin; protease, serine, 8; Prss8; Serine protease 8

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.

2 Citations: Showing 1 - 2
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  1. 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;8(8):e1002937.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. 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;42(8):676-83.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay

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Fluorogenic Peptide Substrates

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