Recombinant Human PGPEP-1 Protein, CF

Catalog # Availability Size / Price Qty
6278-CY-010
Product Details
Citations (1)
FAQs
Reviews

Recombinant Human PGPEP-1 Protein, CF Summary

Purity
>90%, 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 the hydrolysis of pyroglutamyl-AMC. The specific activity is >9,500 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived human PGPEP-1 protein
Met1-His209, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Met1
Predicted Molecular Mass
24 kDa
SDS-PAGE
24-26 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.

6278-CY

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol and DTT.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 5 mM DTT, pH 9.0
  • Recombinant Human PGPEP-1 (rhPGPEP-1) (Catalog # 6278-CY)
  • Substrate: L-pyroglutamic acid-AMC (Pyr-AMC) (Bachem, Catalog # I-1300), 10 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 rhPGPEP-1 to 0.1 ng/μL in Assay Buffer.
2.  Dilute Substrate to 100 μM in Assay Buffer.
3.  Load into a plate 50 μL of 0.1 ng/μL rhPGPEP-1, and start the reaction by adding 50 μL of 100 μM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 100 μM Substrate.
4.  Read plate at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes.
5.  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 7-Amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

Per Well:
  • rhPGPEP-1: 0.005 μg
  • Substrate: 50 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: PGPEP-1

Pyroglutamyl peptidase-1 (PGPEP-1) is an omega peptidase which removes pyroglutamyl residues from the amino termini of peptides and proteins (1). It is a cytosolic cysteine peptidase that is expressed in most cell types (2). The enzyme requires a thiol‑reducing agent for activity (3). PGPEP-1 is potentially involved in the inactivation of biologically active peptides that possess an amino terminal pyroglutamyl group (3). Examples of such peptides include neurotensin, luteinizing hormone releasing hormone, and thyrotropin-releasing hormone.

References
  1. Kilbane Z. et al. (2007) Mol. Cell. Biochem. 297:189.
  2. Cummins P.M. and B. O'Connor (1998) Biochim. Biophys. Acta. 1429:1.
  3. Dando P.M. et al. (2003) Protein Express. Purif. 28:111.
Long Name
Pyroglutamyl Aminopeptidase I
Entrez Gene IDs
54858 (Human)
Alternate Names
EC 3.4.19.3; PAP-I; Pcp; PGPEP1; PGPEP-1; PGPIFLJ20208; PGP-IPAP-I; PGPMGC10812; Pyroglutamyl aminopeptidase I; pyroglutamyl-peptidase 1; pyroglutamyl-peptidase I5-oxoprolyl-peptidase; Pyrrolidone-carboxylate peptidase

Citation for Recombinant Human PGPEP-1 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. Disturbed Ca2+ homeostasis increases glutaminyl cyclase expression; connecting two early pathogenic events in Alzheimer's disease in vitro.
    Authors: De Kimpe L, Bennis A, Zwart R, van Haastert E, Hoozemans J, Scheper W
    PLoS ONE, 2012;7(9):e44674.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Enzyme Assay

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