Recombinant Human Pepsinogen C/PGC Protein, CF

Catalog # Availability Size / Price Qty
6186-AS-010
Product Details
Citations (1)
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Recombinant Human Pepsinogen C/PGC Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005). The specific activity is >700 pmol/min/μg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Pepsinogen C/PGC protein
Met1-Ala388, with a C-terminal 6-His tag
Accession # P20142
Accession #
N-terminal Sequence
Analysis
Ala17 & Leu43
Predicted Molecular Mass
41 kDa & 38 kDa
SDS-PAGE
35-40 kDa, reducing conditions

Product Datasheets

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.

6186-AS

Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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 opening.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Sodium Citrate, 250 mM NaCl, pH 2.5
  • Recombinant Human Pepsinogen C/PGC (rhPGC) (Catalog # 6186-AS)
  • Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(Dnp)-OH (Catalog # ES005), 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 rhPGC to 100 µg/mL in Assay Buffer.
  2. Incubate 100 µg/mL rhPGC for 10 minutes at room temperature.
  3. Dilute 100 µg/mL rhPGC to 0.5 µg/mL in Assay Buffer.
  4. Dilute Substrate to 20 µM in Assay Buffer.
  5. Load 50 µL of 0.5 µg/mL of rhPGC into a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 20 µM Substrate.
  6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     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:
  • rhPGC: 0.025 µg
  • Substrate: 10 µM
Reconstitution Calculator

Reconstitution Calculator

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: Pepsinogen C/PGC

Pepsinogen C (progastricsin) is an aspartic proteinase zymogen that belongs to the peptidase family A1. Pespsin C, the activated protease, is a digestive enzyme that is produced in the stomach and constitutes a major component of the gastric mucosa. This protein is synthesized as an inactive zymogen that is converted into its active mature form at low pH by sequential cleavage of the prosegment in an autocatalytic process (1). Polymorphisms in this gene are associated with susceptibility to gastric cancers (2). Serum levels of this enzyme are used as a biomarker for certain gastric diseases including Helicobacter pylori related gastritis (3).

References
  1. Richter, C. et al. (1998) Biochem. J. 335:481.
  2. Liu, H. J. et al. (2003) World J. Gastroenterol. 9:50.
  3. Plebani, M. (1993) Crit. Rev. Clin. Lab. Sci. 30:273.
Alternate Names
EC 3.4.23; EC 3.4.23.3; FLJ99563; Gastricsin; PEPC; pepsin C; Pepsinogen C; pepsinogen group II; PGC; PGII; preprogastricsin; progastricsin (pepsinogen C)

Citation for Recombinant Human Pepsinogen C/PGC 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. Global protease activity profiling provides differential diagnosis of pancreatic cysts
    Authors: SL Ivry, JM Sharib, DA Dominguez, N Roy, SE Hatcher, M Yip-Schnei, CM Schmidt, RE Brand, WG Park, M Hebrok, G Kim, AJ O'Donoghue, KS Kirkwood, CS Craik
    Clin. Cancer Res., 2017;0(0):.
    Applications: Bioassay

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Fluorogenic Peptide Substrates

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