Carboxypeptidase B1, encoded by the CPB1 gene, specifically cleaves the C-terminal Arg and Lys residues with a preference for Arg (1). Expressed mainly in pancreas, CPB1 is a useful serum marker for acute pancreatitis (2). The deduced amino acid sequence of human CPB1 consists of a signal peptide (residues 1 to 15), a pro region (residue 16 to 110), and a mature chain (residues 111 to 417). The purified rhCPB1 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).
Recombinant Human Carboxypeptidase B1/CPB1 Protein, CF
R&D Systems | Catalog # 2897-ZN
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Carboxypeptidase B1/CPB1 Protein (2897-ZN)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
NS0
Accession Number
Structure / Form
Pro form
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Carboxypeptidase B1/CPB1 protein
His16-Tyr417, with a C-terminal 10-His tag
His16-Tyr417, with a C-terminal 10-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
His16
Predicted Molecular Mass
47 kDa
SDS-PAGE
45 kDa, reducing conditions
Activity
Measured by its ability to cleave a colorimetric peptide substrate, Hippuryl-Arg.
The specific activity is >1,000 pmol/min/µg, as measured under the described conditions.
The specific activity is >1,000 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2897-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.
|
Background: Carboxypeptidase B1/CPB1
References
- Aviles, F.X. and J. Vendrell (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) p. 831, Academic Press, San Diego.
- Yamamoto, K.K. et al. (1992) J. Biol. Chem. 267:2575.
Alternate Names
CPB1, PASP, PCPB
Gene Symbol
CPB1
UniProt
Additional Carboxypeptidase B1/CPB1 Products
Product Documents for Recombinant Human Carboxypeptidase B1/CPB1 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Carboxypeptidase B1/CPB1 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Carboxypeptidase B1/CPB1 Protein, CF
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Protocols
View specific protocols for Recombinant Human Carboxypeptidase B1/CPB1 Protein, CF (2897-ZN):
Materials
- Activation Buffer: 50 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% Brij-35, pH 7.5 (TCNB)
- Assay Buffer: 50 mM Tris, 100 mM NaCl, pH 7.5
- Recombinant Human Carboxypeptidase B1/CPB1 (rhCPB1) (Catalog # 2897-ZN)
- Trypsin (Sigma, Catalog # T-1426)
- Substrate: Hippuryl-R (Sigma, Catalog # H-2508), 25 mM in diH2O
- 96 well clear UV-transparent microplate (Corning, Catalog # 3635)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhCPB1 to 100 μg/mL with 1.0 μg/mL Trypsin in Activation Buffer.
- Incubate at room temperature for 30 minutes.
- Dilute active rhCPB1 to 2 μg/mL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- Load 50 μL of the 2 μg/mL rhCPB1 into a plate. Include a Substrate blank with 50 μL of Assay Buffer.
- Start the reaction by adding 50 μL of diluted 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).
- rhCPB1: 0.1 μg
- Substrate: 1 mM
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