Recombinant Human Pepsinogen A Protein, CF

R&D Systems | Catalog # 6155-AS

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Pepsinogen A Protein (6155-AS)
  • 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

P00790

Applications

Enzyme Activity
View all Pepsinogen A Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Pepsinogen A protein
Met1-Ala388, with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ile16 and Val63

Predicted Molecular Mass

41 kDa

SDS-PAGE

40-43 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >7,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6155-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.

Background: Pepsinogen A

Pepsins are aspartic proteases that are synthesized in the gastric mucosa and secreted into the stomach. They are released as zymogens called pepsinogens and then converted to active pepsins by the acidic pH of gastric juices (1). PGA-3, PGA-4, and PGA-5 are isozymogens of human Pepsinogen A, which differ in amino acid sequence by 2-4 residues (2). This recombinant human Pepsinogen A corresponds to PGA-4. Pepsins have optimal activity under conditions of acidic pH and are inhibited by pepstatin. Pepsin A has broad substrate specificity, but preferentially cleaves peptide bonds involving aromatic and aliphatic amino acids.

References

  1. Athauda, S.B. et al. (1989) J. Biochem. 106:920.
  2. Zwiers, A. et al. (1994) Clin. Nephrol. 41:153.

Alternate Names

EC 3.4.23, EC 3.4.23.1, FLJ58952, FLJ77962, pepsin A, pepsinogen 4, group I (pepsinogen A), pepsinogen A4, PGA3, PGA5

Entrez Gene IDs

643847 (Human)

Gene Symbol

PGA4

UniProt

P00790

Additional Pepsinogen A Products

Product Documents for Recombinant Human Pepsinogen A Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human Pepsinogen A Protein, CF

For research use only

Related Research Areas

Aspartic Proteases and Regulators

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Protocols

View specific protocols for Recombinant Human Pepsinogen A Protein, CF (6155-AS):

Materials
  • Activation Buffer: 50 mM Sodium Citrate, pH 2.5
  • Assay Buffer: 50 mM Sodium Citrate, 500 mM NaCl, pH 4.0
  • Recombinant Human Pepsinogen A (rhPepsinogen A) (Catalog # 6155-AS)
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002), 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 rhPepsinogen A to 100 µg/mL in Activation Buffer.
  2. Incubate for 15 minutes at room temperature.
  3. Dilute activated rhPepsinogen A to 0.02 µg/mL in Assay Buffer.
  4. Dilute Substrate to 20 µM in Assay Buffer.
  5. Load 50 µL of 0.02 µg/mL of rhPepsinogen A 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:

  • rhPepsinogen A: 0.001 µg
  • Substrate: 10 µM

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