Recombinant Human Pro-Collagen II Protein, CF

Catalog #: 7589-CL
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
7589-CL-020

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Citations (1)

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Pro-Collagen II Protein, CF Summary

Product Specifications

Purity

>85%, 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 II 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 II protein
Gln26-Arg266 & Gly957-Leu1487, with an N-terminal 5-His tag

Accession #

P02458

N-terminal Sequence
Analysis

His & Asp1242

Predicted Molecular Mass

78 kDa

SDS-PAGE

92-100 kDa, 65-70 kDa, 36 kDa, reducing conditions

Product Datasheets

7589-CL

Product Datasheet

COA

SDS

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.

7589-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 II (rhCOL-2A1) (Catalog # 7589-CL)
Recombinant Human BMP‑1/PCP (rhBMP-1) (Catalog # 1927-ZN)
Reducing SDS-PAGE Sample Buffer
SDS-PAGE or Western Blot

Dilute rhCOL-2A1 to 75 μg/mL in Dilution Buffer.
Dilute rhBMP-1 to 10 μg/mL in Assay Buffer.
Combine one volume of diluted rhCOL-2A1 with three volumes of diluted rhBMP-1. For controls, combine one volume of rhCOL-2A1 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 one hour.
After incubation, combine rhCOL-2A1/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 rhCOL-2A1) per lane and analyze the cleavage by SDS-PAGE followed by protein staining and/or Western blot.
Activity calculation:

% Cleavage = [1 -	% full-length rhCOL-2A1 (reaction)	] x 100%
	% full-length rhCOL-2A1 (control)

Per Lane:

rhCOL-2A1: 0.5 µg
rhBMP-1: 0.2 µg

Reconstitution Calculator

Background: Collagen II

Type II collagen is an essential component of cartilage. It is essential for skeletal development and for the ability of cartilage to resist compressive forces. Type II collagen is a homotrimer of a1(II) chains. Defects in Type II collagen are the cause of numerous diseases, including chondrodysplasias and arthritis (1). This recombinant mini pro-a1(II) collagen consists of the N‑terminal propeptide, 108 Gly-X-Y repeats, and the C‑terminal propeptide. Proteolytic processing of the C‑terminal propeptide by the procollagen C-proteinase (BMP-1) releases chondrocalcin, a calcium binding protein.

References

Kuivaniemi H. et al. (1997) Hum. Mutat. 9:300.

Entrez Gene IDs

1280 (Human); 12824 (Mouse); 25412 (Rat)

Alternate Names

Alpha-1 type II collagen; ANFH; AOM; cartilage collagen; Chondrocalcin; COL11A3; COL2A1; Collagen 2; collagen alpha-1(II) chain; Collagen II; collagen II, alpha-1 polypeptide; collagen type 2; collagen, type II, alpha 1 (primary osteoarthritis, spondyloepiphysealdysplasia, congenital); collagen, type II, alpha 1; Collagen-2; MGC131516; SEDC

Related Research Areas

Citation for Recombinant Human Pro-Collagen II 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.

1 Citation: Showing 1 - 1

A Highly Sensitive Biomarker of Type II Collagen C-Terminal Pro-Peptide Associated with Cartilage Formation
Authors: H Port, AC Bay-Jensen, Y He, MA Karsdal, T Gantzel, CS Thudium, S Holm Niels
International Journal of Molecular Sciences, 2022-12-27;24(1):.
Species: Human
Sample Types: Whole Cells
Applications: Cell Culture