Recombinant Human NPEPPS Protein, CF
Recombinant Human NPEPPS Protein, CF Summary
Product Specifications
Pro46-Val919, with an N-terminal Met and a C-terminal 10-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.
6410-ZN
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol. |
| 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: 25 mM HEPES, 1 mM DTT, pH 7.0
- Recombinant Human Puromycin‑sensitive aminopeptidase/NPEPPS (rhNPEPPS) (Catalog # 6410-ZN)
- Substrate: Leu-AMC (Bachem, Catalog # I-1240), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhNPEPPS to 0.2 ng/μL in Assay Buffer.
- Dilute Substrate to 20 μM in Assay Buffer.
- Load 50 μL of 0.2 ng/μL rhNPEPPS into the microplate and start the reaction by adding 50 μL of Substrate. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively, for 5 minutes in kinetic mode.
- 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.
- rhNPEPPS: 0.010 µg
- Substrate: 10 µM
Reconstitution Calculator
Background: Puromycin-sensitive aminopeptidase/NPEPPS
NPEPPS, also known as Puromycin-sensitive aminopeptidase and cytosol alanyl aminopeptidase, is a zinc metallopeptidase which hydrolyzes N‑terminal amino acids from its substrates. NPEPPS is expressed in most tissues as a cytoplasmic protein, but a membrane-associated form has been identified in the brain (1, 2). The enzyme has broad substrate specificity but prefers basic and hydrophobic residues in the substrate P1 and P'1 sites (3). NPEPPS is known to hydrolyze dynorphins and enkephalins, and may contribute to the degradation of these peptides in the brain (4). NPEPPS plays a major role in the degradation of toxic polyglutaminyl peptides that are released by proteasomes (5). NPEPPS is also known to degrade the tau protein, which accumulates and polymerizes in some neurodegenerative diseases (6).
- Dyer, S.H. et al. (1990) J. Neurochem. 54:547.
- McLellan, S. et al. (1988) J. Neurochem. 51:1552.
- Johnson, G.D. and L.B. Hersh (1990) Arch. Biochem. Biophys. 276:305.
- Safavi, A. and L.B. Hersh (1995) J. Neurochem. 65:389.
- Bhutani, N. et al. (2007) EMBO J. 26:1385.
- Sengupta, S. et al. (2006) Biochemistry 45:15111.
Product Specific Notices
Coomassie is a registered trademark of Imperial Chemical Industries Ltd.Citations for Recombinant Human NPEPPS 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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Rapid imaging of lung cancer using a red fluorescent probe to detect dipeptidyl peptidase 4 and puromycin-sensitive aminopeptidase activities
Authors: S Kawashima, D Yoshida, T Yoshioka, A Ogasawara, K Fujita, M Yanagiya, M Nagano, C Konoeda, H Hino, K Kitano, M Sato, R Hino, R Kojima, T Komatsu, M Kamiya, Y Urano, J Nakajima
Scientific Reports, 2022-06-01;12(1):9100.
Species: Human
Sample Types: Recombinant Proteins
Applications: Bioassay -
Proteasome inhibitors alter levels of intracellular peptides in HEK293T and SH-SY5Y cells.
Authors: Dasgupta S, Castro L, Dulman R, Yang C, Schmidt M, Ferro E, Fricker L
PLoS ONE, 2014-07-31;9(7):e103604.
Species: Human
Sample Types: Protein
Applications: Bioassay
FAQs
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