Recombinant Human PGPEP-1 Protein, CF Summary
Product Specifications
Met1-His209, 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.
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.
|
Assay Procedure
- 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).
- rhPGPEP-1: 0.005 μg
- Substrate: 50 μM
Reconstitution Calculator
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.
- Kilbane Z. et al. (2007) Mol. Cell. Biochem. 297:189.
- Cummins P.M. and B. O'Connor (1998) Biochim. Biophys. Acta. 1429:1.
- Dando P.M. et al. (2003) Protein Express. Purif. 28:111.
Product Specific Notices
Coomassie is a registered trademark of Imperial Chemical Industries Ltd.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
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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, 2012-09-07;7(9):e44674.
Species: Human
Sample Types: Cell Lysates
Applications: Enzyme Assay
FAQs
-
Do you have a unit conversion for pmol/min/μg to units?
Conversion to International Units would require testing our enzyme directly side-by-side with a biological reference material from NIBSC. The NIBSC catalog does not currently have any reference materials for the PGPEP-1 protein, so this is not currently possible.
Additionally, the assay to measure activity of this protein involves reading fluorescence of the fluorogenic peptide substrate in kinetic mode. Since we're reading in kinetic mode, we're specifically interested in the change in fluorescence over time. That means that time is an integral part of the pmol/min/μg units.
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