Recombinant Mouse Marapsin/Pancreasin Protein, CF

Catalog # Availability Size / Price Qty
1989-SE-010

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Recombinant Mouse Marapsin/Pancreasin Protein, CF Summary

Product Specifications

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.
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.
Source
Mouse myeloma cell line, NS0-derived mouse Marapsin/Pancreasin protein
Ala23-Thr290, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Ala23 & Met38
Predicted Molecular Mass
28 kDa
SDS-PAGE
33 kDa, reducing conditions

Product Datasheets

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1989-SE

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.

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.

Assay Procedure

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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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.
Entrez Gene IDs
83886 (Human); 213171 (Mouse)
Alternate Names
CAP-2; CAPH2; channel-activating protease 2; EC 3.4.21; EC 3.4.21.-; EC 3.4.21.4; EC 3.4.21.43; Marapsin; MPN; MPNMarapsin; Pancreasin; protease, serine 27; PRSS27; serine protease 27

Citation for Recombinant Mouse Marapsin/Pancreasin 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.

1 Citation: Showing 1 - 1

  1. Mutational tail loss is an evolutionary mechanism for liberating marapsins and other type I serine proteases from transmembrane anchors.
    Authors: Raman K, Trivedi N, Raymond W, Ganesan R, Kirchhofer D, Verghese G, Craik C, Schneider E, Nimishakavi S, Caughey G
    J Biol Chem, 2013-02-27;288(15):10588-98.
    Species: Mouse
    Sample Types: Serum
    Applications: Bioassay

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