Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF

R&D Systems | Catalog # 6220-CL

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein (6220-CL)
  • 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

CHO

Accession Number

P02452

Applications

Enzyme Activity
View all Collagen I alpha 1 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Collagen I alpha 1 protein
Gln23-Lys277 + Gly1094-Leu1464, with an N-terminal 6-His tag.

Purity

>80%, 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

His & Asp1219

Predicted Molecular Mass

65 kDa (full-length), 40 kDa, 28 kDa

SDS-PAGE

79-83 kDa, 54-56 kDa, 35-36 kDa, reducing conditions

Activity

Measured by the cleavage of its C-terminal propeptide by Recombinant Human BMP‑1/PCP (Catalog # 1927-ZN).
>50% of full-length Pro-Collagen I alpha 1 is cleaved by Recombinant Human BMP‑1/PCP (Catalog # 1927-ZN), as measured under the described conditions.

Formulation, Preparation, and Storage

6220-CL
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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: Collagen I alpha 1

Type I collagen is the most abundant structural protein of connective tissues such as skin, bone and tendon. It is synthesized as a procollagen molecule which is characterized by a 300 nm triple helical domain flanked by globular N- and C-terminal propeptides (1). The triple helical domain contains Gly-Xaa-Yaa triplets where Xaa and Yaa are frequently proline and hydroxyproline, respectively. The non-helical propeptides are removed by procollagen N- and C-proteinase activities so that the mature triple helices can self-assemble into collagen fibrils that provide tensile strength to tissues (1). Type I collagen is a heterotrimer that consists of two alpha 1(I) chains and one alpha 2(I) chain, although homotrimers consisting of three identical alpha 1(I) chains have also been described (2). This recombinant mini pro-alpha 1(I) collagen consists of a shortened alpha 1(I) chain with following domain structure from N- to C-terminus: N-propeptide, N‑telopeptide, the 33 most N-terminal Gly-Xaa-Yaa repeats, the 33 most C-terminal Gly-Xaa-Yaa repeats, C-telopeptide and C-propeptide. The preparation contains a mixture of the full-length molecule, pN collagen I( alpha 1) and the C-terminal propeptide. This truncated pro-alpha 1(I) collagen is a substrate for procollagen N-proteinase and procollagen C-proteinase.

References

  1. Canty, E.G. et al. (2005) J. Cell Sci. 118:1341.
  2. Han, S. et al. (2008) J. Mol. Biol. 383:122.

Alternate Names

COL1A1, OI4

Entrez Gene IDs

1277 (Human); 12842 (Mouse); 29393 (Rat)

Gene Symbol

COL1A1

UniProt

P02452

Additional Collagen I alpha 1 Products

Product Documents for Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF

For research use only

Citations for Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF

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Protocols

View specific protocols for Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF (6220-CL):

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 Pro-Collagen I alpha 1 (rhPro-COL1A1) (Catalog # 6220-CL)
  • Recombinant Human BMP‑1/PCP (BMP‑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 rhBMP-1 to 5 µg/mL in Assay Buffer.
  3. Combine one volume of diluted rhPro-COL1A1 with three volumes of diluted rhBMP-1. For controls, combine one volume of rhPro‑COL1A1 with three volumes of Assay Buffer, as well as, three volumes of rhBMP-1 with one volume of Assay Buffer.
  4. Incubate reaction vials and controls at 37 °C for 1 hour.
  5. After incubation, combine rhPro-COL1A1/rhBMP-1 reaction mixtures and controls with reducing SDS-PAGE gel buffer at a 2:1 (reaction mixture:gel buffer) ratio (v/v).  Mix and incubate samples at 95-100 °C for 3-5 minutes to stop reactions.
  6. 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.
  7. Activity Calculation:
     % Cleavage = [1  -   % full-length rhPro-COL1A1 (reaction) ] x 100%
% full-length rhPro-COL1A1 (control)

Per Lane:

  • rhPro-COL1A1: 0.5 µg
  • rhBMP-1: 0.1 µg

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