Carboxypeptidase A1 encoded by the CPA1 gene cleaves the C-terminal amide or ester bond of peptides that have a free C-terminal carboxyl group (1). It prefers the C-terminal residues with aromatic or branched aliphatic side chains including Phe, Tyr, Trp, Leu or Ile. It is important in the degradation of food proteins to produce essential amino acids such as Phe and Trp. The deduced amino acid sequence of mouse CPA1 consists of a signal peptide (residues 1 to 16), a pro region (residue 17 to 110), and a mature chain (residues 111 to 419). The purified recombinant CPA1 corresponds to the pro form, which can be activated and assayed under the conditions described in the Activity Assay Protocol.
Recombinant Mouse Carboxypeptidase A1/CPA1 Protein, CF
R&D Systems | Catalog # 2765-ZN
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Carboxypeptidase A1/CPA1 Protein (2765-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 mouse Carboxypeptidase A1/CPA1 protein
Asn17-Tyr419, with a C-terminal 10-His tag
Asn17-Tyr419, with a C-terminal 10-His tag
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Asn17
Predicted Molecular Mass
47 kDa
SDS-PAGE
42 kDa, reducing conditions
Activity
Measured by its ability to cleave the colorimetric peptide substrate Ac-Phe-Thiaphe-OH in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB). Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468.
The specific activity is >2,500 pmol/min/µg, as measured under the described conditions.
The specific activity is >2,500 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2765-ZN
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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.
|
Background: Carboxypeptidase A1/CPA1
References
- Auld, D.S. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) p. 812 Academic Press, San Diego.
Alternate Names
CPA1
Gene Symbol
CPA1
UniProt
Additional Carboxypeptidase A1/CPA1 Products
Product Documents for Recombinant Mouse Carboxypeptidase A1/CPA1 Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse Carboxypeptidase A1/CPA1 Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse Carboxypeptidase A1/CPA1 Protein, CF (2765-ZN):
Materials
*Adjusted for Substrate Blank
**Using the extinction coefficient 13260 M-1cm-1
***Using the path correction 0.320 cm
Note: the output of many spectrophotometers is in mOD. Per Well:
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Recombinant Mouse Carboxypeptidase A1/CPA1 (rmCPA1) (Catalog # 2765-ZN)
- Trypsin (Sigma, Catalog # T-1426)
- Substrate: Ac-Phe-Thiaphe-OH (Peptides International, Catalog # STP-3621-PI), 10 mM stock in DMSO
- 5,5’-dithio-bis(2-nitrobenzoic acid) (DTNB), (Sigma, Catalog # D-8130), 10 mM stock in DMSO
- 96 well clear plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rmCPA1 to 100 µg/mL with 1.0 µg/mL Trypsin in Assay Buffer.
- Incubate at room temperature for 60 minutes.
- Dilute active rmCPA1 to 0.2 µg/mL in Assay Buffer.
- Combine equal volumes of 10 mM Substrate and 10 mM DTNB. Then, dilute this mixture to 200 µM Substrate, 200 µM DTNB with Assay Buffer.
- Load 50 µL of the 0.2 µg/mL rmCPA1 into a plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
- Read in kinetic mode for 5 minutes at an absorbance of 405 nm.
- Calculate specific activity using the following formula:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol |
| ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Using the extinction coefficient 13260 M-1cm-1
***Using the path correction 0.320 cm
Note: the output of many spectrophotometers is in mOD. Per Well:
- rmCPA1: 0.01 μg
- Substrate: 100 µM
- DTNB: 100 µM
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