Catalog #: 6220-CL
7 Citations Datasheet / COA / SDS
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas
Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF Summary
Product Specifications
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-CollagenI alpha 1is cleaved byRecombinant 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.
|
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-CollagenI 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
- Dilute rhPro-COL1A1 to 75 µg/mL in Dilution Buffer.
- Dilute rhBMP-1 to 5 µg/mL in Assay Buffer.
- 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.
- Incubate reaction vials and controls at 37 °C for 1 hour.
- 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.
- 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) |
Per Lane:
- rhPro-COL1A1: 0.5 µg
- rhBMP-1: 0.1 µg
Reconstitution Calculator
Reconstitution Calculator
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) collagenconsists 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.
- Canty, E.G. et al. (2005) J. Cell Sci. 118:1341.
- 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
Related Research Areas
- Extracellular Matrix Molecules
- Molecules Secreted by VSMC
- Osteogenesis Markers
- Osteoprogenitor Markers
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.
7 Citations: Showing 1 - 7
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- Cancer immunotherapy by NC410, a LAIR-2 Fc protein blocking human LAIR-collagen interaction
Authors: MIP Ramos, L Tian, EJ de Ruiter, C Song, A Paucarmayt, A Singh, E Elshof, SV Vijver, J Shaik, J Bosiacki, Z Cusumano, C Jensen, N Willumsen, MA Karsdal, L Liu, S Langermann, S Willems, D Flies, L Meyaard
Elife, 2021-06-14;10(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay - Integrins and extracellular matrix proteins modulate adipocyte thermogenic capacity
Authors: MA Gonzalez P, K Stojkova, MK Vaicik, A Pelowe, A Goddi, A Carmona, B Long, AA Qutub, A Gonzalez, RN Cohen, EM Brey
Scientific Reports, 2021-03-08;11(1):5442.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay - Development of a miniaturized 96-Transwell air-liquid interface human small airway epithelial model
Authors: T Bluhmki, S Bitzer, JA Gindele, E Schruf, T Kiechle, M Webster, J Schymeinsk, R Ries, F Gantner, D Bischoff, J Garnett, R Heilker
Sci Rep, 2020-08-03;10(1):13022.
Species: Human
Sample Types: Cell Culture Supernates, Whole Cells
Applications: Bioassay, ELISA Capture - 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, 2019-07-18;9(1):10414.
Species: Human
Sample Types: Whole Cells
Applications: Cell Culture - HIV-1 Tat Regulates Occludin and A? Transfer Receptor Expression in Brain Endothelial Cells via Rho/ROCK Signaling Pathway
Oxid Med Cell Longev, 2016-08-02;2016(0):4196572.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay - 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, 2014-05-30;8(2):507-512.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay - 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, 2013-02-25;8(2):e57160.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay
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