|"Protein lysates of osteosarcoma cell lines were used in this study. The lysates were prepared in the same way as in our previous study. A lysate of HuO9 cells was used to examine the gel images of individual samples, and lysates of HuO9, HOS, MG-63, MNNG-HOS, 14 3B, HS-Os-1, NOS-1, and NOS-10 cells were used to create the common internal standard sample employed in this study. In brief, the monolayer cells were washed twice with ice-cold PBS, and fixed with 10% tri chloro acetic acid for 30 min. The cells were then scraped off, and washed with PBS. They were then treated with protein lysis buffer (2M thiourea, 6M urea, 3% CHAPS, 1% Triton X-100) for 30 min. After centrifugation at 15,000 rpm for 30 min, the supernatant was recovered as a protein lysate.
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Citation: Tani Y, Tajima T, Kawai A, Unuma Y , Kinoshita H, et al. (2014) Evaluation of a Novel Automated Machine, the Auto2D, for Two-Dimensional Gel Electrophoresis. J Proteomics Bioinform 7: 108-111."