Recombinant Human Hepsin Protein, CF

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R&D Systems Recombinant Proteins and Enzymes
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Citations (5)
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Recombinant Human Hepsin Protein, CF Summary

Product Specifications

>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.
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.
Mouse myeloma cell line, NS0-derived human Hepsin protein
Arg45-Leu417 (Asp161Glu & Arg162Lys), with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Predicted Molecular Mass
42 kDa
40-43 kDa, reducing conditions

Product Datasheets

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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.


Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

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
Reconstitution Calculator

Reconstitution Calculator

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Background: Hepsin

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.
Entrez Gene IDs
3249 (Human); 15451 (Mouse); 29135 (Rat)
Alternate Names
EC 3.4.21; EC; 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

Citations for Recombinant Human Hepsin 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.

5 Citations: Showing 1 - 5
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  1. Anti-Tumor Functions of Prelatent Antithrombin on Glioblastoma Multiforme Cells
    Authors: J Peñas-Mart, G Luengo-Gil, S Espín, N Bohdan, C Ortega-Sab, MC Ródenas, D Zaragoza-H, MJ López-Andr, C Plasencia, V Vicente, A Carmona-Ba, I Martínez-M
    Biomedicines, 2021;9(5):.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  2. Structure-activity relationship studies of dipeptide-based hepsin inhibitors with Arg bioisosteres
    Authors: H Kwon, H Ha, H Jeon, J Jang, SH Son, K Lee, SK Park, Y Byun
    Bioorganic chemistry, 2020;0(0):104521.
    Species: N/A
    Sample Types: Peptide
    Applications: Bioassay
  3. Design and synthesis of dye-conjugated hepsin inhibitors
    Authors: K Kim, H Kwon, D Choi, T Lim, I Minn, SH Son, Y Byun
    Bioorg. Chem., 2019;89(0):102990.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  4. Design, Synthesis, and Testing of Potent, Selective Hepsin Inhibitors via Application of an Automated Closed-Loop Optimization Platform.
    Authors: Pant S, Mukonoweshuro A, Desai B, Ramjee M, Selway C, Tarver G, Wright A, Birchall K, Chapman T, Tervonen T, Klefstrom J
    J Med Chem, 2018;61(10):4335-4347.
    Applications: Enzyme Assay
  5. Deregulated hepsin protease activity confers oncogenicity by concomitantly augmenting HGF/MET signalling and disrupting epithelial cohesion.
    Authors: Tervonen T, Belitskin D, Pant S, Englund J, Marques E, Ala-Hongisto H, Nevalaita L, Sihto H, Heikkila P, Leidenius M, Hewitson K, Ramachandra M, Moilanen A, Joensuu H, Kovanen P, Poso A, Klefstrom J
    Oncogene, 2015;0(0):ePub.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay


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Recombinant Human Hepsin Protein, CF
By Anonymous on 05/22/2019
Application: Immunoassay Standard

AF4776 was used as the detection antibody in a sandwich ELISA with MAB4776 as the capture antibody and 4776-SE-010 as the calibrator.

Recombinant Human Hepsin Protein, CF
By Anonymous on 04/20/2019
Application: Enzymatic activity in vitro

Activated hepsin was used at a concentration of 0.03nM. Assay was performed in PBS. The hepsin inhibitor was from Pant et al., 2018 (J Med Chem. 2018 May 24;61(10):4335-4347. doi: 10.1021/acs.jmedchem.7b01698. Epub 2018 May 14.
Design, Synthesis, and Testing of Potent, Selective Hepsin Inhibitors via Application of an Automated Closed-Loop Optimization Platform.)

Recombinant Human Hepsin Protein, CF
By Anonymous on 04/20/2019
Application: Silver stained gel to test purity

Loaded 0.1ug, reduces SDS-PAGE on gradient gel, followed by silver staining