Recombinant Mouse Carboxypeptidase B1/CPB1 Protein, CF
Recombinant Mouse Carboxypeptidase B1/CPB1 Protein, CF Summary
Product Specifications
Ser16-Tyr415, with a C-terminal 10-His tag
Accession # NP_083982
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.
2898-ZN
| 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
- Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Assay Buffer: 25 mM Tris, 0.1 M NaCl, pH 7.5
- Recombinant Mouse Carboxypeptidase B1/CPB1 (rmCPB1) (Catalog # 2898-ZN)
- Trypsin (Sigma, Catalog # T-1426)
- AEBSF (Catalog # EI001), 100 mM stock in DMSO
- Substrate: Hippuryl-Arg (Sigma, Catalog # H-2508), 25 mM in diH2O
- 96-well Clear UV Plate (Corning, Catalog # 3635)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Activate rmCPB1 at 100 µg/mL with 1 µg/mL Trypsin in Activation Buffer.
- Incubate at room temperature for 30 minutes.
- Add AEBSF at a final concentration of 1 mM to stop reaction.
- Dilute activated rmCPB1 to 2 µg/mL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- In a plate, load 50 µL of 2 µg/mL rmCPB1, and include a Substrate Blank containing 50 µL Assay Buffer.
- Start the reaction by adding 50 µL of 2 mM Substrate into wells.
- Read in kinetic mode for 5 minutes at an absorbance of 254 nm.
- Calculate specific activity using the following formula:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (OD/min) x Conversion Factor** (pmol/OD) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard Hippuric acid (Sigma Catalog # 112003).
- rmCPB1: 0.1 µg
- Substrate: 1 mM
Reconstitution Calculator
Background: Carboxypeptidase B1/CPB1
Carboxypeptidase B1, encoded by the CPB1 gene, specifically cleaves the C-terminal Arg and Lys residues with a preference for Arg (1). The deduced amino acid sequence of mouse CPB1 consists of a signal peptide (residues 1 to 15), a pro region (residue 16 to 108), and a mature chain (residues 109 to 415). The purified rmCPB1 corresponds to the pro form, which can be activated by trypsin, the only pancreatic protease capable of generating active enzyme from the zymogen in vitro (1).
- Aviles, F.X. and J. Vendrell (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 831, Academic Press, San Diego.
Citation for Recombinant Mouse Carboxypeptidase B1/CPB1 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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Site-specific carboxypeptidase B1 tyrosine nitration and pathophysiological implications following its physical association with nitric oxide synthase-3 in experimental sepsis.
Authors: Chatterjee S, Lardinois O, Bonini MG, Bhattacharjee S, Stadler K, Corbett J, Deterding LJ, Tomer KB, Kadiiska M, Mason RP
J. Immunol., 2009-08-28;183(6):4055-66.
Applications: ELISA (Standard)
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