Recombinant Human QPCT Protein, CF
Recombinant Human QPCT Protein, CF Summary
Product Specifications
Ala33-Leu361, with an N-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.
6368-ZN
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol. |
| 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: 25 mM HEPES, pH 7.0
- rhPGPEP-1 Buffer: 0.1 M Tris, 5 mM DTT, pH 9.0
- Recombinant Human Glutaminyl‑peptide Cyclotransferase/QPCT (rhQPCT) (Catalog # 6368-ZN)
- Substrate: Q-AMC (Bachem, Catalog # I-1175)
- Recombinant Human PGPEP-1 (rhPGPEP-1) (Catalog # 6278-CY)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhQPCT to 0.2 μg/mL in Assay Buffer.
- Dilute Substrate to 200 μM in Assay Buffer.
- Combine 150 μL of 0.2 ug/mL rhQPCT and 150 μL of 200 μM Substrate. Include a Substrate Blank containing 150 μL Assay Buffer and 150 μL of 200 μM Substrate.
- Incubate reaction at room temperature for 20 minutes.
- Stop reaction by boiling at 100 °C for 5 minutes and cool on ice for 3 minutes.
- Dilute rhPGPEP-1 to 1 μg/mL in rhPGPEP-1 Buffer.
- Add 300 μL of 1 μg/mL rhPGPEP-1 to each vial.
- Incubate reaction at room temperature for 10 minutes.
- Load 100 μL in quadruplicate from each vial into a 96 well plate.
- Read plate at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in endpoint mode.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU) |
| Incubation time (min) x amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard 7-Amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).
- rhQPCT: 0.005 μg
- Substrate: 50 μM
Reconstitution Calculator
Background: Glutaminyl-peptide Cyclotransferase/QPCT
Glutaminyl-peptide Cyclotransferase, also known as Glutaminyl Cyclase, catalyzes the conversion of N‑terminal L-glutaminyl residues of peptides to pyroglutamyl groups (1). The enzyme is present in the pituitary and adrenal glands, where it is important for the generation of the N‑terminal pyroglutamyl groups of peptide hormones such as neurotensin and thyrotropin-releasing hormone. Glutaminyl Cyclase also catalyzes the conversion of N‑terminal L‑glutamyl residues to pyroglutamyl residues (2). This activity may contribute to the formation of several amyloid-related plaque forming peptides, contributing to Alzheimer’s disease pathology. Glutaminyl Cyclase is also considered to be a diagnostic marker of thyroid tumors (3).
- Busby, W.H. Jr. et al. (1987) J. Biol. Chem. 262:8532.
- Schilling, S. et al. (2004) FEBS Lett. 563:191.
- Griffith, O.L. et al. (2006) J. Clin. Oncol. 24:5043.
Citations for Recombinant Human QPCT 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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Citations: Showing 1 - 2
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Discovery of potent indazole-based human glutaminyl cyclase (QC) inhibitors as Anti-Alzheimer's disease agents
Authors: N Van Manh, VH Hoang, VTH Ngo, S Kang, JJ Jeong, HJ Ha, H Kim, YH Kim, J Ann, J Lee
European Journal of Medicinal Chemistry, 2022-10-12;244(0):114837.
Species: Human
Sample Types:
Applications: Bioassay -
Discovery of highly potent human glutaminyl cyclase (QC) inhibitors as anti-Alzheimer's agents by the combination of pharmacophore-based and structure-based design
Authors: N Van Manh, VH Hoang, VTH Ngo, J Ann, TH Jang, JH Ha, JY Song, HJ Ha, H Kim, YH Kim, J Lee, J Lee
European Journal of Medicinal Chemistry, 2021-09-08;226(0):113819.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay
FAQs
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What Isoform is present in Recombinant Human QPCT Protein, CF (Catalog # 6368-ZN-010)?
The Isoform present in Recombinant Human QPCT Protein, CF (Catalog # 6368-ZN-010) is Isoform 1.
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