spring leaf collagen 6 in 1.5% (w/v) in a solution of 1% Tween 20 for 30 min at room temperature. The collagen was then washed with PBS and dried over MgSO 4. The resulting powder was purified by SDS-PAGE and eluted with water.
The peptide was elute with a 1:1 mixture of Triton X-100 (Sigma-Aldrich) and 1 mM EDTA (pH 7.4) at 4°C for 1 h. After washing with TBS, the eluate was dried with 1× Saffron-X (Pierce) for 20 min. A final elution of the peptides was performed with 0.1% Sulfuric Acid (sulfate buffer) (1:100) to remove the sulfate. This eluting step was repeated with the remaining peptidoglycan. Protein was separated by electrophoresis on a Bio-Rad Electron Microscopy System (Bio-RAD) with an Agilent 2100 (Agilence) column (Thermo Fisher Scientific). The eluent was washed twice with S-buffered saline (TBS) before being elutinated with 2× TFA (0.2% w/w) containing 0% NaCl. Finally, a final 1 min elutation was conducted with 3× NaOH (50 mM Tris-HCl, pH 7) followed by a 5 min incubation at 37° C. for 5 h at which time the protein was removed by centrifugation. For the determination of protein content, protein concentration was determined by using a Masson-Cox protein assay kit (Masson).
, and the following values were calculated using the formula:
. These values are expressed as the mean ± SEM of three independent experiments.
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spring leaf collagen review
. J. Clin. Invest. 118 : 689-697 View in Article Scopus (0)
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The company’s website says it has been in business since 1892 and has a history of producing high-quality products. The company has also been involved in the production of high quality products for the pharmaceutical industry.
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