详细说明
- 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.
- ActivityMeasured 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. See Activity Assay Protocol on www.RnDSystems.com
- SourceMouse myeloma cell line, NS0-derived Met1-Ala468, with a C-terminal 10-His tag
- Accession #
- N-terminal Sequence
AnalysisNo result, Gln25 predicted. - Predicted Molecular Mass49 kDa
- SDS-PAGE52-57 kDa, reducing conditions
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.
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- 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 # )
- Recombinant Human BMP‑1/PCP (rhBMP-1) (Catalog # )
- Reducing SDS-PAGE Sample Buffer
- SDS-PAGE or Western Blot
- Dilute rhPro-COL1A1 to 75 µg/mL in Dilution Buffer.
- Dilute rmPCPE-1 to 15 µg/mL in Assay Buffer.
- 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).
- Incubate reaction vials and controls at 37 °C for 30 minutes.
- Dilute rhBMP-1 to 1.875 µg/mL in Assay Buffer.
- 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.
- Incubate reaction vials and controls at 37 °C for 30 minutes.
- 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.
- 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.
- Activity calculation:
% Cleavage = [1 - | % full-length rhPro-COL1A1 (reaction) | ] x 100% |
% full-length rhPro-COL1A1 (control) |
- rmPCPE-1: 0.1 µg
- rhPro-COL1A1: 0.5 µg
- rhBMP-1: 0.0125 µg
- References:
- Steiglitz, B.M. et al. (2002) J. Biol. Chem. 277:49820.
- Kessler, E. et al. (1990) Biochem. Biophys. Res. Commun. 173:81.
- Moali, C. et al. (2005) J. Biol. Chem. 280:24188.
- 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