Recombinant Human PRCP Protein, CF
Recombinant Human PRCP Protein, CF Summary
Product Specifications
Met1-His496, 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.
7164-SE
| Formulation | Supplied as a 0.2 μm filtered solution in MES 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.
|
Assay Procedure
- Assay Buffer: 50 mM Sodium Acetate, 0.1 M NaCl, pH 4.0
- Recombinant Human Lysosomal Pro‑X Carboxypeptidase/PRCP (rhPRCP) (Catalog # 7164-SE)
- Substrate: Z-Pro-Ala (Bachem, Catalog # C-2485), 100 mM stock in deionized water
- 2-Mercaptoethanol (Sigma, Catalog # M7154)
- NaOH, 2 M stock in deionized water
- o-Phthaldialdehyde (OPA) (Sigma, Catalog # P0657), 0.373 M in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhPRCP to 0.2 µg/mL in Assay Buffer.
- Dilute Substrate to 400 µM in Assay Buffer.
- Mix 125 μL of 0.2 µg/mL rhPRCP and 125 μL 400 µM Substrate for a final concentration of 0.1 µg/mL and 200 µM respectively. Include a control containing 125 μL of 0.2 µg/mL rhPRCP that has been heat treated for 5 minutes at 100 °C and 125 μL 400 µM Substrate.
- Incubate for 30 minutes at 37 °C.
- Stop reaction by adding 250 μL of a solution containing 15 mM o-PA in 0.2 M NaOH containing 0.1% (v/v) 2-Mercaptoethanol.
- Incubate for 10 minutes at room temperature.
- Load 200 µL of the incubated samples in duplicate into the plate.
- Read at excitation and emission wavelengths of 330 nm and 450 nm (top read), 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 heat treated control
**Derived using calibration standard L-Alanine (Sigma, Catalog # A7469).
- rhPRCP: 0.01 µg
- Substrate: 0.1 mM
Reconstitution Calculator
Background: Lysosomal Pro-X Carboxypeptidase/PRCP
Lysosomal Pro-X Carboxypeptidase (PRCP) is a serine carboxypeptidase found principally in lysosomes, but can also be secreted or associated with the plasma membrane. The enzyme is specific for peptide substrates with a penultimate proline residue. PRCP is also known as Angiotensinase C because of its activity against angiotensin II and angiotensin III (1). PRCP is also known to be involved in the activation of prekallikrein on the surface of endothelial cells (2). Because of these activities, PRCP is considered to be a cardioprotective enzyme (3). PRCP has been shown to degrade the bioactive peptide a-melonocyte stimulating hormone, thereby destroying its activity (4). Therefore PRCP may play a role in the melanocortin signaling pathway and the regulation of energy metabolism (5).
- Yang, H.Y.T. et al. (1968) Nature 218:1224.
- Shariat-Madar, Z. et al. (2002) J. Biol. Chem. 277:17962.
- Mallela, J. et al. (2009) Int. J. Biochem. Cell Biol. 41:477.
- Wallingford, N. et al. (2009) J. Clin. Invest. 119:2291.
- Diano, S. et al. (2011) Front. Neuroendocrinol. 32:70.
Citation for Recombinant Human PRCP 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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Fibroblast Activation Protein Cleaves and Inactivates Fibroblast Growth Factor 21
Authors: DR Dunshee, TW Bainbridge, NM Kljavin, J Zavala-Sol, AC Schroeder, R Chan, R Corpuz, M Wong, W Zhou, G Deshmukh, J Ly, DP Sutherlin, JA Ernst, J Sonoda
J. Biol. Chem, 2016-01-21;291(11):5986-96.
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