Recombinant Human Hepsin Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Hepsin Protein, CF Summary

Details of Functionality
Measured by its ability to cleave tert-butoxycarbonyl-Gln-Arg-Arg-7-amino-4-methylcoumarin (Boc-QRR-AMC). The specific activity is >20,000 pmol/min/μg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Hepsin protein
Arg45-Leu417 (Asp161Glu & Arg162Lys), with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Arg45
Protein/Peptide Type
Recombinant Enzymes
Gene
HPN
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
42 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
40-43 kDa, reducing conditions
Publications
Read Publications using
4776-SE in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Activation Buffer: 0.1 M Tris, 10 mM CaCl2, 0.15 M NaCl, 0.05 % Brij-35, pH 8.0
  • Assay Buffer: 50 mM Tris, pH 9.0
  • Recombinant Human Hepsin (rhHepsin) (Catalog # 4776-SE)
  • Fluorogenic Peptide Substrate: BOC-Gln-Arg-Arg-AMC (Bachem, Catalog # I-1655), 5 mM stock in 50:50 DMSO:Methanol
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhHepsin to 100 µg/mL in Activation Buffer.
  2. Incubate at 37 °C for 24 hours.
  3. Dilute activated rhHepsin to 0.2 ng/µL in Assay Buffer.
  4. Dilute Substrate to 200 µM in Assay Buffer.
  5. Load 50 µL of the 0.2 ng/µL rhHepsin in a black well plate and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate.
  6. Read at excitation and emission wavelengths of 380 nm and 460 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 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891).

Per Well:
  • rhHepsin: 0.010 µg
  • Substrate: 100 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Hepsin Protein, CF

  • EC 3.4.21
  • EC 3.4.21.106
  • Hepsin
  • HPN
  • serine protease hepsin catalytic chain
  • serine protease hepsin
  • TMPRSS1
  • TMPRSS1serine protease hepsin non-catalytic chain
  • Transmembrane protease serine 1
  • transmembrane protease, serine 1

Background

Hepsin, also known as TMPRSS1, is a Type II membrane protein with an extracellular serine protease domain (1). It is most highly expressed in liver, but is also present in many other tissues, notably lung, kidney, and skeletal muscle (2). A soluble form of Hepsin lacking the transmembrane domain has been identified (3). Hepsin is capable of activating Factor VII, and may initiate blood coagulation at the cell surface (4). Hepsin is overexpressed in various human tumors, including prostate (5), and is considered to be a biomarker for prostate cancer (6). Recombinant human Hepsin was expressed as a secreted, soluble protein lacking its cytosolic and transmembrane domains. The D161E and R162K mutations were introduced into the prosequence to improve expression of the recombinant human Hepsin.

  1. Leytus, S.P. et al. (1988) Biochemistry 27:1067.
  2. Tsuji, A. et al. (1991) J. Biol. Chem. 266:16948.
  3. Li, Y. et al. (2005) Biomed. Biochim. Acta 1681:157.
  4. Kazama, Y. et al. (1995) J. Biol. Chem. 270:66.
  5. Dhanasekaran, S.M. et al. (2001) Nature 412:822.
  6. Wu, Q. and Parry, G. (2007) Front. Biosci. 12:5052.

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Publications for Hepsin (4776-SE)(3)

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Bioinformatics

Gene Symbol HPN
Uniprot