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

Catalog # Availability Size / Price Qty
6220-CL-020
Product Details
Citations (4)
FAQs
Supplemental Products
Reviews

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

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.
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.
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.
Accession #
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

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.

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.

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 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
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.

=
÷

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.
Entrez Gene IDs
1277 (Human)
Alternate Names
Alpha-1 type I collagen; COL1A1; COL-IA1; Collagen 1 alpha 1; Collagen 1; collagen alpha 1 chain type I; collagen alpha-1(I) chain; Collagen I alpha 1; collagen of skin, tendon and bone, alpha-1 chain; Collagen Type I Alpha 1 Chain; collagen, type I, alpha 1; Collagen1; Collagen-1; EDSARTH1; OI4; pro-alpha-1 collagen type 1; Type I Procollagen Alpha 1 Chain

Citations for Recombinant Human Pro-Collagen I alpha 1/COL1A1 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.

4 Citations: Showing 1 - 4
Filter your results:

Filter by:

  1. Endothelial tip cells in vitro are less glycolytic and have a more flexible response to metabolic stress than non-tip cells
    Authors: B Yetkin-Ari, IMC Vogels, N Neyazi, V van Duinen, RH Houtkooper, CJF van Noorde, I Klaassen, RO Schlingema
    Sci Rep, 2019;9(1):10414.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  2. HIV-1 Tat Regulates Occludin and A? Transfer Receptor Expression in Brain Endothelial Cells via Rho/ROCK Signaling Pathway
    Oxid Med Cell Longev, 2016;2016(0):4196572.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. Role of secreted type I collagen derived from stromal cells in two breast cancer cell lines.
    Authors: Kim, Sung Hoo, Lee, Hye Yoon, Jung, Seung Pi, Kim, Sangmin, Lee, Jeong Eo, Nam, Seok Jin, Bae, Jeoung W
    Oncol Lett, 2014;8(2):507-512.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Nicotinamide inhibits vasculogenic mimicry, an alternative vascularization pathway observed in highly aggressive melanoma.
    Authors: Itzhaki, Orit, Greenberg, Eyal, Shalmon, Bruria, Kubi, Adva, Treves, Avraham, Shapira-Frommer, Ronnie, Avivi, Camilla, Ortenberg, Rona, Ben-Ami, Eytan, Schachter, Jacob, Besser, Michal J, Markel, Gal
    PLoS ONE, 2013;8(2):e57160.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

FAQs

No product specific FAQs exist for this product, however you may

View all Proteins and Enzyme FAQs

Recombinant Enzymes

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

There are currently no reviews for this product. Be the first to review Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF and earn rewards!

Have you used Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review