Recombinant Human PCPE-1 Protein, CF

Catalog # Availability Size / Price Qty
2627-PE-020
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Recombinant Human PCPE-1 Protein, CF Summary

Purity
>95%, 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 Human PCPE-1, is processed by BMP-1, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human PCPE-1 protein
Met1-Asp449, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
No result, Gln26 expected
Predicted Molecular Mass
47 kDa
SDS-PAGE
51-57 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.

2627-PE

Formulation Supplied as a 0.2 μm filtered solution in MES 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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -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 Human PCPE-1 (rhPCPE-1) (Catalog # 2627-PE)
  • Recombinant Human Pro Collagen I  alpha 1 (rhCOL1A1) (Catalog # 6220-CL)
  • Recombinant Human BMP‑1/PCP (rhBMP-1) (Catalog # 1927-ZN)
  • Reducing SDS-PAGE Sample Buffer (3X or greater)
  • SDS-PAGE or Western Blot
  1. Dilute rhCOL1A1 to 75 µg/mL in Dilution Buffer.
  2. Dilute rhPCPE-1 to 15 µg/mL in Assay Buffer.
  3. In a tube, combine equal volumes (a minimum of 15 µL of each is required for this assay) of diluted rhCOL1A1 and diluted rhPCPE‑1. Prepare samples in duplicate (one replicate to be used as a control).
  4. Incubate vials at 37 °C for 30 minutes.
  5. Dilute rhBMP-1 to 1.875 µg/mL in Assay Buffer.
  6. Add one equivalent (1/2 the total volume of rhCOL1A1/rhPCPE-1 mixtures) of diluted rhBMP-1 to each reaction tube. Prevent the reaction from occurring in the control duplicates by immediately mixing with reducing gel buffer (see step 8 for volumes).
  7. Incubate reaction tubes at 37 °C for 30 minutes.
  8. After incubation, combine 40 µL of each reaction mixture and control with 20 µL of reducing SDS-PAGE gel buffer. Mix and incubate samples at 95-100 °C for 3-5 minutes.
  9. Load 30 µL per lane and analyze the cleavage by SDS-PAGE followed by protein staining and/or Western blot.
  10. Activity calculation:
     % Cleavage = [1 -  % full-length rhCOL1A1 (reaction) ] x 100%
% full-length rhCOL1A1 (control)
Per Lane:
  • rhPCPE-1: 0.1 µg
  • rhCOL1A1: 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 and 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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