Recombinant Mouse HAI-1 Protein, CF

R&D Systems | Catalog # 1141-PI

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse HAI-1 Protein (1141-PI)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

NP_058603

Applications

Inhibition Activity
View all HAI-1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse HAI-1 protein
Glu30-Glu443 (Cys325Ser), with a C-terminal 10-His tag

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.

N-terminal Sequence Analysis

Glu30

Predicted Molecular Mass

48 kDa

SDS-PAGE

60 kDa, reducing conditions

Activity

Measured by its ability to inhibit trypsin cleavage of a fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The IC50 value is <1.5 nM, as measured under the described conditions.

Formulation, Preparation, and Storage

1141-PI
Formulation Lyophilized from a 0.2 μm filtered solution in Tris, NaCl and CaCl2.
Reconstitution

Reconstitute at 100 μg/mL in sterile, deionized water.
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 reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷

Background: HAI-1

Hepatocyte growth factor activator inhibitor-1 (HAI-1) encoded by the Spint1 gene is a Kunitz-type serine protease inhibitor, identified as a strong inhibitor of HGF activator (HGFA) and matripase (1). The membrane-anchored HAI-1 consists of two Kunitz domains, a LDL-receptor-like domain, and a C-terminal transmembrane domain (2). Two soluble forms are generated by ectodomain shedding, one with a single Kunitz domain and the other with two Kunitz domains. HAI-1 is not only an inhibitor but also a specific receptor of active HGFA, acting as a reservoir of this enzyme on the cell surface (3). The shedding of HAI-1 and HGFA/HAI-1 complex is enhanced by treatment with phorbol 12-myristate, 13-acetate or IL-1 beta. The regulated shedding is completely inhibited by a synthetic zinc metalloprotease inhibitor (3).

References

  1. Denda, et al. (2002) J. Biol. Chem. 277:14053.
  2. Shimomura, et al. (1997) J. Biol. Chem. 272:6370.
  3. Kataoka, et al. (2000) J. Biol. Chem. 275:40453.

Long Name

HGF Activator Inhibitor Type 1

Alternate Names

HAI1, SPINT1

Entrez Gene IDs

6692 (Human); 20732 (Mouse)

Gene Symbol

SPINT1

UniProt

NP_058603

Additional HAI-1 Products

Product Documents for Recombinant Mouse HAI-1 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Mouse HAI-1 Protein, CF

For research use only

Related Research Areas

Serine Proteases and Regulators

Citations for Recombinant Mouse HAI-1 Protein, CF

Customer Reviews for Recombinant Mouse HAI-1 Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Mouse HAI-1 Protein, CF and earn rewards!

Have you used Recombinant Mouse HAI-1 Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Mouse HAI-1 Protein, CF (1141-PI):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Mouse HAI-1 (rmHAI-1) (Catalog # 1141-PI)
  • Trypsin (Sigma, Catalog # T-1426)
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute Trypsin to 0.25 µg/mL with Assay Buffer.
  2. Prepare a curve of rmHAI-1 (MW: 47792 Da) in Assay Buffer. Make the following serial dilutions: 100, 40, 20, 12, 8, 4, 2 and 1 nM.
  3. Mix equal volumes of the rmHAI-1 curve dilutions and the diluted Trypsin. Include a Trypsin control (in duplicate) containing Assay Buffer and the diluted Trypsin without any rmHAI-1.
  4. Incubate reactions for 60 minutes at 37 °C.
  5. After incubation, dilute the mixtures five-fold in Assay Buffer.
  6. Dilute Substrate to 20 µM in Assay Buffer.
  7. Load into plate 50 µL of the diluted incubated mixtures, and start the reaction by adding 50 µL of 20 µM Substrate.
  8. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  9. Calculate specific activity for each point using the following formula (if needed):

     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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:

  • Trypsin: 0.00125 µg
  • rmHAI-1: 5 nM, 2 nM, 1 nM, 0.6 nM, 0.4 nM, 0.2 nM, 0.1 nM, 0.05 nM
  • Substrate: 10 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes