Recombinant Human Pepsinogen C/PGC Protein, CF Summary
Product Specifications
Met1-Ala388, with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
|
Assay Procedure
- 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
- Dilute rhPGC to 100 µg/mL in Assay Buffer.
- Incubate 100 µg/mL rhPGC for 10 minutes at room temperature.
- Dilute 100 µg/mL rhPGC to 0.5 µg/mL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- 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.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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).
- rhPGC: 0.025 µg
- Substrate: 10 µM
Reconstitution Calculator
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).
- Richter, C. et al. (1998) Biochem. J. 335:481.
- Liu, H. J. et al. (2003) World J. Gastroenterol. 9:50.
- Plebani, M. (1993) Crit. Rev. Clin. Lab. Sci. 30:273.
Citations 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.
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A Prototype Assay Multiplexing SARS-CoV-2 3CL-Protease and Angiotensin-Converting Enzyme 2 for Saliva-Based Diagnostics in COVID-19
Authors: Suresh, V;Sheik, DA;Detomasi, TC;Zhao, T;Zepeda, T;Saladi, S;Rajesh, UC;Byers, K;Craik, CS;Davisson, VJ;
Biosensors
Applications: Bioassay -
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., 2017-04-19;0(0):.
Applications: Bioassay
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