Recombinant Mouse PCPE-1 Protein, CF

Catalog # Availability Size / Price Qty
2239-PE-020
Product Details
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Recombinant Mouse PCPE-1 Protein, CF Summary

Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by the stimulation of procollagen type I processing by BMP-1. >50% of the full-length procollagen, in the presence of recombinant mouse PCPE, is processed by BMP-1, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived mouse PCPE-1 protein
Met1-Ala468, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
No result, Gln25 predicted.
Predicted Molecular Mass
49 kDa
SDS-PAGE
52-57 kDa, reducing conditions

Product Datasheets

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.

2239-PE

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
  • Dilution Buffer: 50 mM Tris, 5 mM CaCl2, pH 7.5
  • Assay Buffer: 50 mM Tris, 5 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
  • Recombinant Mouse PCPE‑1 (rmPCPE-1) (Catalog # 2239-PE)
  • Recombinant Human Pro-Collagen I  alpha 1 (rhPro‑COL1A1) (Catalog # 6220-CL)
  • Recombinant Human BMP‑1/PCP (rhBMP-1) (Catalog # 1927-ZN)
  • Reducing SDS-PAGE Sample Buffer
  • SDS-PAGE or Western Blot
  1. Dilute rhPro-COL1A1 to 75 µg/mL in Dilution Buffer.
  2. Dilute rmPCPE-1 to 15 µg/mL in Assay Buffer.
  3. Combine 20 µL of diluted rhPro-COL1A1 and 20 µL of diluted rmPCPE-1. Prepare a rhPro-COL1A1 control by combining 20 µL of rhPro‑COL1A1 and 20 µL of Assay Buffer. Prepare one rmPCPE-1 control by combining 20 µL of rmPCPE-1 and 20 µL of Assay Buffer (optional).
  4. Incubate reaction vials and controls at 37 °C for 30 minutes.
  5. Dilute rhBMP-1 to 1.875 µg/mL in Assay Buffer.
  6. Add 20 µL of diluted rhBMP-1 to each reaction vial, excluding the rhPro-COL1A1 control. Add 20 µL of Assay Buffer to the rhPro‑COL1A1 control in place of rhBMP-1.
  7. Incubate reaction vials and controls at 37 °C for 30 minutes.
  8. After incubation, combine 30 µL of each reaction mixture and control with 30 µL of reducing SDS-PAGE gel buffer. Mix and incubate samples at 95-100 °C for 3-5 minutes to stop reactions.
  9. Load 40 µL (0.5 µg of rhPro-COL1A1) per lane and analyze the cleavage by SDS-PAGE followed by protein staining and/or Western blot.
  10. Activity calculation:
     % Cleavage = [1 -   % full-length rhPro-COL1A1 (reaction) ] x 100%
% full-length rhPro-COL1A1 (control)
Per Lane
  • rmPCPE-1: 0.1 µg
  • rhPro-COL1A1: 0.5 µg
  • rhBMP-1: 0.0125 µg
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: PCPE-1

Procollagen C-endopeptidase enhancers, known as PCPEs or PCOLCEs, are secreted extracellular matrix glycoproteins that consist of two CUB domains and one NTR domain. They are known to stimulate enzymatic cleavage of procollagens I‑III by the BMP-1/tolloid family of metalloproteases, also known as procollagen C‑proteinases (1). PCPE‑1 is expressed primarily by interstitial connective tissues such as tendons, calvaria, and skin (2). Although BMP-1/tolloid proteinases are involved in processing of multiple extracellular proteins, the enhancer activity of PCPE-1 is specific to procollagens since it has no effect on BMP‑1/tolloid cleavage of other substrates (3). It is thought that PCPE-1 enhances cleavage of procollagens by binding to the substrate, inducing a conformation change in the substrate (3), although interaction between PCPE-1 and full-length BMP‑1/tolloid proteinases has also been reported (4).

References
  1. Steiglitz, B.M. et al. (2002) J. Biol. Chem. 277:49820.
  2. Kessler, E. et al. (1990) Biochem. Biophys. Res. Commun. 173:81.
  3. Moali, C. et al. (2005) J. Biol. Chem. 280:24188.
  4. Ge, G. et al. (2006) J. Biol. Chem. 281:10786.
Long Name
Procollagen C-Proteinase Enhancer-1
Entrez Gene IDs
5118 (Human); 18542 (Mouse)
Alternate Names
PCOLCE; PCPE; PCPE1; PCPE-1; PCPE1Type I procollagen COOH-terminal proteinase enhancer; procollagen C-endopeptidase enhancer 1; procollagen C-endopeptidase enhancer; Procollagen COOH-terminal proteinase enhancer 1; Procollagen C-proteinase enhancer 1; procollagen, type 1, COOH-terminal proteinase enhancer; Type 1 procollagen C-proteinase enhancer protein

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