Recombinant Human HAI-2A Protein, CF

Images

 
There are currently no images for HAI-2A (1106-PI).

Every product we sell is backed by Novus' 100% Guarantee. If you have used this product, please submit your images and reviews to earn reward points.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Inhibition Activity
Format
Carrier-Free

Order Details

Recombinant Human HAI-2A Protein, CF Summary

Details of Functionality
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 <4.6 nM, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human HAI-2A protein
Met1-Lys197, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Ala28
Protein/Peptide Type
Recombinant Enzymes
Gene
SPINT2
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
21 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
27 kDa and 32 kDa, reducing conditions
Publications
Read Publications using
1106-PI 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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in Tris, NaCl and CaCl2.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 100 μg/mL in sterile, deionized water.
Assay Procedure
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Human HAI-2A (rhHAI-2A) (Catalog # 1106-PI)
  • Trypsin (Sigma, Catalog # T-1426)
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002) ), 2 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 Trypsin to 0.25 µg/mL with Assay Buffer.
  2. Prepare a curve of rhHAI-2A (MW: 20539 Da) in Assay Buffer. Make the following serial dilutions: 1000, 100, 33.3, 16.7, 8.33, 4.17, 2.08, and 0.417 nM.
  3. Mix equal volumes of the rhHAI-2A curve dilutions and the diluted Trypsin. Include a control (in duplicate) containing Assay Buffer and the diluted Trypsin without any rhHAI-2A.
  4. Incubate reactions for 30 minutes at room temperature.
  5. After incubation, dilute the mixtures by 5 fold in Assay Buffer.
  6. Dilute Substrate to 20 µM in Assay Buffer.
  7. Load 50 µL of the diluted incubated mixtures into a plate, 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. Derive the 50% inhibition concentration (IC50) by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.
  10. The specific activity for Trypsin at each point may be determined 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
  • rhHAI-2A curve: 50, 5, 1.67, 0.83, 0.417, 0.209, 0.104, and 0.0209 nM
  • Substrate: 10 µM

Notes

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

Alternate Names for Recombinant Human HAI-2A Protein, CF

  • HAI2A
  • HAI-2A
  • Placental Bikunin
  • SPINT2

Background

Two alternatively spliced forms of HAI-2 have been found in human tissues (1). HAI-2A, the full-length molecule and also known as placental bikunin, is a major form expressed in human tissues. Encoded by the SPINT2 gene, HAI-2A consists of two Kunitz domains, and a C-terminal transmembrane domain (1‑5). Both Kunitz domains can function as inhibitors independent of each other. In addition to HGF activator and trysin, HAI-2A strongly inhibits plasmin, tissue and plasma kallikreins, and factor XIa. In comparison, HAI-2A is a weaker inhibitor of factor VIIa-tissue factor, factors IXa, Xa, and XIIa. Recombinant HAI-2A prolonged the clotting time in an activated partial thromboplastin time assay.

  1. Kataoka, H. et al. (2002) Biochem. Biophys. Res. Comm. 290:1096.
  2. Kawaguchi, T. et al. (1997) J. Biol. Chem. 272:27558.
  3. Marlor. C.W. et al. (1997) J. Biol. Chem. 272:12202.
  4. Muller-Pillasch, F. et al. (1998) Biochim. Biophys. Acta 1395:88.
  5. Delaria, K.A. et al. (1997) J. Biol. Chem. 272:12209.

Customers Who Viewed This Item Also Viewed...

NBP2-45041
Species: Hu, Po
Applications: Flow, ICC/IF, IF
NB100-524
Species: Hu, Mu
Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, Flow-IC
NBP1-85466
Species: Hu, Mu, Fe
Applications: WB, IHC, IHC-P
NBP2-67605
Species: Hu, Mu
Applications: WB, ICC/IF, IHC, IHC-P, IP
NBP1-88220
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P, KD
NB100-1598
Species: Hu, Mu
Applications: WB, Flow, ICC/IF
NBP2-98675
Species: Hu
Applications: WB, IHC, IHC-P, IP
NB600-534
Species: Hu, Mu, Po, V-Vi
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, In vitro, RIA
NBP2-15895
Species: Hu, Mu, Rt
Applications: WB, IHC, IHC-P
NB600-1071
Species: Mu, Rt
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP
AF3846
Species: Hu, Mu, Rt
Applications: WB, IHC
NBP2-44520
Species: Hu, Ca(-), Rt(-)
Applications: WB, Flow, ICC/IF, IHC, IHC-P, Flow-IC, IF, MI
NBP1-87581
Species: Hu
Applications: WB, IHC, IHC-P
NBP2-20127
Species: Hu, Mu
Applications: WB, ICC/IF, IHC, IHC-P, IP

Publications for HAI-2A (1106-PI)(3)

Reviews for HAI-2A (1106-PI) (0)

There are no reviews for HAI-2A (1106-PI). By submitting a review you will receive an Amazon e-Gift Card or Novus Product Discount.
  • Review with no image -- $10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen
  • Review with an image -- $25/€18/£15/$25 CAD/¥150 Yuan/¥2500 Yen

FAQs for HAI-2A (1106-PI) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Customers Who Bought This Also Bought

Contact Information

Product PDFs

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Review this Product

Be the first to review our Recombinant Human HAI-2A Protein, CF and receive a gift card or discount.

Bioinformatics

Gene Symbol SPINT2
Uniprot