Recombinant Mouse Carboxypeptidase B1/CPB1 Protein, CF

Catalog #: 2898-ZN
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
2898-ZN-010

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse Carboxypeptidase B1/CPB1 Protein, CF Summary

Product Specifications

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.

Activity

Measured by its ability to cleave a colorimetric peptide substrate, Hippuryl-Arg. The specific activity is >5,000 pmol/min/µg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived mouse Carboxypeptidase B1/CPB1 protein
Ser16-Tyr415, with a C-terminal 10-His tag
Accession # NP_083982

Accession #

NP_083982

N-terminal Sequence
Analysis

Ser16

Predicted Molecular Mass

47 kDa

SDS-PAGE

43 kDa and 48 kDa, reducing conditions

Product Datasheets

2898-ZN

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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. 6 months from date of receipt, -20 to -70 °C as supplied. 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials

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 V_max* (OD/min) x Conversion Factor** (pmol/OD)
	amount of enzyme (µg)

*Adjusted for Substrate Blank
**Derived using calibration standard Hippuric acid (Sigma Catalog # 112003).

Per Well:

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).

References

Aviles, F.X. and J. Vendrell (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 831, Academic Press, San Diego.

Entrez Gene IDs

1360 (Human); 76703 (Mouse)

Alternate Names

carboxypeptidase B; carboxypeptidase B1 (tissue); Carboxypeptidase B1; CPB; CPB1; DKFZp779K1333; EC 3.4.17; EC 3.4.17.2; Pancreas-specific protein; PASP; PCPB; procarboxypeptidase B

Related Research Areas

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.

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