Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF Summary
Details of Functionality
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.
>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.
Applications/Dilutions
Dilutions
Enzyme Activity
Theoretical MW
65 kDa (full-length), 40 kDa, 28 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Purity
>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
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
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
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human Pro-Collagen I alpha 1/COL1A1 Protein, CF
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
Background
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.
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FAQs for Collagen I alpha 1 (6220-CL). (Showing 1 - 4 of 4 FAQs).
We are looking for an anti-Collagen antibody for Flow Cytometry (FACS) analysis in Swine sample. Would you please provide some references if there is any available as well?
We currently have one antibody for porcine Collagen validated in flow cytometry (catalog # NBP1-94202).
I am looking for anti collagen 1 antibody, I see your product calls it collagen I alpha 1 is there a difference?
Collagen I alpha 1 is the most common protein for the collagen 1 protein. There are other subunits as well (such as collagen 1 alpha 2…etc.).
The predicted molecular weight says 150kDA but the band on the gel appears at 130kDA why is that?
Differences between the predicted and observed MW can be affected by a number of factors including PTMs, relative charges and experimental factors. These cumulative effect of these differences can be quite substantial with larger molecules such as collagen I alpha 1.
How long does delivery take for your collagen 1 alpha 1 antibody?
Our most popular collagen I alpha 1 antibody (NB600-408) is usually in stock and ships priority overnight for next business day delivery in the U.S.
The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.
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