Recombinant Mouse Marapsin/Pancreasin Protein, CF

R&D Systems | Catalog # 1989-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Marapsin/Pancreasin Protein (1989-SE)
  • 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

Q8BJR6

Applications

Enzyme Activity
View all Marapsin/Pancreasin Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Marapsin/Pancreasin protein
Ala23-Thr290, with a C-terminal 6-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

Ala23 & Met38

Predicted Molecular Mass

28 kDa

SDS-PAGE

33 kDa, reducing conditions

Activity

Measured by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Gly-Arg-ThioBenzyl ester (Z-GR-SBzl), in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB). Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468.
The specific activity is >1,200 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

1989-SE
Formulation Lyophilized from a 0.2 μm filtered solution in MES and NaCl.
Reconstitution Reconstitute at 100 μg/mL in sterile 25 mM MES and 100 mM NaCl, pH 6.5.
Shipping The product is shipped at ambient temperature. 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.

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Background: Marapsin/Pancreasin

Marapsin, Pancreasin, and channel-activating protease 2 (CAP-2), encoded by the Prss27 gene, are different names given for the same serine protease that is expressed strongly in the pancreas (1). The mouse protein is synthesized with a signal peptide (amino acid residues 1‑22), a pro peptide (residues 23‑37) and a mature chain (residues 38‑290) corresponding to the serine protease domain. The full-length protein was expressed and the secreted protein purified. The N-terminal sequencing results indicate that the purified protein is a disulfide bond-linked dimer formed between the pro peptide and the mature chain. The active enzyme has low activity against peptide substrates tested, but high activity against thioester substrates. The peptidase activity is inhibited by 20 mM benzamidine.

References

  1. Bhagwandin, V.J. et al. (2003) J. Biol. Chem. 278:3363.

Alternate Names

CAP-2, MPN, Pancreasin, PRSS27

Entrez Gene IDs

83886 (Human); 213171 (Mouse)

Gene Symbol

PRSS27

UniProt

Q8BJR6

Additional Marapsin/Pancreasin Products

Product Documents for Recombinant Mouse Marapsin/Pancreasin Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Mouse Marapsin/Pancreasin Protein, CF

For research use only

Related Research Areas

Serine Proteases and Regulators

Citations for Recombinant Mouse Marapsin/Pancreasin Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Marapsin/Pancreasin Protein, CF (1989-SE):

Materials
  • Assay Buffer: 50 mM Tris, 250 mM NaCl, 0.05% (v/v) Brij-35, pH 8.0
  • Recombinant Mouse Marapsin/Pancreasin (rmMPN) (Catalog # 1989-SE)
  • Substrate: Z-Gly-Arg-SBzl (MP Biomedicals, Catalog # SB007)
  • 5,5’Dithio-bis(2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130)
  • 96 well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rmMPN to 1.0 ng/µL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer with 200 µM of DTNB.
  3. Load 50 µL of the diluted rmMPN in the plate, and start the reaction by adding 50 µL of the Substrate/DTNB mixture to wells. Include a Substrate Blank containing Assay Buffer and Substrate mix.
  4. Read in kinetic mode for 5 minutes at an absorbance of 405 nm.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank 
     **Using the extinction coefficient 13260 M-1cm-1 
     ***Using the path correction 0.320 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rmMPN: 0.050 µg
  • DTNB: 100 µM
  • Substrate: 100 µM

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