Recombinant Human SPINK1 Protein, CF

Images

 
There are currently no images for SPINK1 (7496-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 Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human SPINK1 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 <10 nM, as measured under the described conditions.
Source
E. coli-derived human SPINK1 protein
Asp24-Cys79, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Met
Protein/Peptide Type
Recombinant Enzymes
Gene
SPINK1
Purity
>85%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
7 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
8-11 kDa, reducing conditions

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 MES, NaCl, CaCl2, DTT, Glycerol and Brij-35.
Purity
>85%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
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 SPINK1 (rhSPINK1) (Catalog # 7496-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 in Assay Buffer.
  2. Prepare a curve of rhSPINK1 (MW: 7201 Da) in Assay Buffer. Make the following serial dilutions: 2000, 1000, 500, 250, 125, 62.5, 31.3, 15.6, 7.81 and 3.91 nM.
  3. Combine equal volumes of 0.25 µg/mL Trypsin with rhSPINK1 serial curve dilutions. Include two controls containing 0.25 µg/mL Trypsin with Assay Buffer.
  4. Incubate reaction mixtures at room temperature for 30 minutes.
  5. After incubation, dilute reaction mixtures 5-fold with Assay Buffer. (Example: 40 µL reaction mixture + 160 µL Assay Buffer).
  6. Dilute Substrate to 20 µM in Assay Buffer.
  7. In a plate, load 50 µL of the diluted reaction mixtures, and start the reaction by adding 50 µL of 20 µM Substrate to wells.
  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% inhibiting concentration (IC50) value for rhSPINK1 by plotting final concentration per well of rhSPINK1 vs. specific activity with 4-PL fitting.
  10. 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-P-L-OH (Bachem, Catalog # M-1975).

  • Trypsin: 0.00125 µg
  • rhSPINK1 curve: 100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.781, 0.391 and 0.195 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 SPINK1 Protein, CF

  • pancreatic secretory trypsin inhibitor
  • PCTT
  • PCTTSpink3
  • PSTI
  • PSTISerine protease inhibitor Kazal-type 1
  • serine peptidase inhibitor, Kazal type 1
  • serine protease inhibitor, Kazal type 1
  • SPINK1
  • Spink3
  • TATI
  • TATITumor-associated trypsin inhibitor

Background

SPINK1 is a serine protease inhibitor with one Kazal-like domain that is secreted  by acinar cells of the pancreas.  It is an inhibitor of trypsin that prevents the premature activation of protease zymogens in the pancreas.  Deficiencies in SPINK1 are a cause of hereditary pancreatitis and tropical calcific pancreatitis (1, 2).  Because elevated SPINK1 expression has been observed with several tumor types, the protein is also known as the tumor-associated trypsin inhibitor (3).
  1. Deybach J.C.D. et al. (2003) Hum. Genet. 113:369.
  2. Chandak G.R. et al. (2002) J. Med. Genet. 39:347.
  3. Huhtala M.L. et al. (1982) J. Biol. Chem. 257:13713.

Customers Who Viewed This Item Also Viewed...

AF3848
Species: Hu
Applications: WB, IHC, IP
MAB25031
Species: Hu
Applications: WB, IHC, IP
H00136541-P01
Species: Hu
NBP2-62664
Species: Hu
Applications: IHC, IHC-P
NBP1-60109
Species: Hu
Applications: WB
MAB3586
Species: Hu
Applications: WB, IP
AF1927
Species: Hu
Applications: WB, IP, ICC
AF7164
Species: Hu
Applications: WB
AF6278
Species: Hu, Mu
Applications: WB
NBP2-12553
Species: Hu
H00008767-M02
Species: Hu, Mu, Rt
Applications: WB, ELISA, IHC-P
NBP2-24503
Species: Hu, Mu, Pm
Applications: WB, ICC/IF, IHC, IHC-P
355-BM/CF
Species: Hu, Mu, Rt
MAB41281
Species: Hu
Applications: WB, Flow, IHC, CyTOF-ready
AF4478
Species: Hu
Applications: WB, IHC

Publications for SPINK1 (7496-PI) (0)

There are no publications for SPINK1 (7496-PI).
By submitting your publication information earn gift cards and discounts for future purchases.

Reviews for SPINK1 (7496-PI) (0)

There are no reviews for SPINK1 (7496-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
  • 1st to review with an image -- $50/€35/£30/$50 CAD/¥300 Yuan/¥5000 Yen

FAQs for SPINK1 (7496-PI) (0)

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

Additional Array Products

Bioinformatics Tool for SPINK1 (7496-PI)

Discover related pathways, diseases and genes to SPINK1 (7496-PI). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for SPINK1 (7496-PI)

Discover more about diseases related to SPINK1 (7496-PI).
 

Pathways for SPINK1 (7496-PI)

View related products by pathway.

PTMs for SPINK1 (7496-PI)

Learn more about PTMs related to SPINK1 (7496-PI).

Blogs on SPINK1

There are no specific blogs for SPINK1, but you can read our latest blog posts.

Customers Who Bought This Also Bought

SPINK1 Antibody (4D4)
H00006690-M01

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 SPINK1 Protein, CF and receive a gift card or discount.

Bioinformatics

Gene Symbol SPINK1
Entrez
Uniprot