Aeg 1

Western blot is widely used in biochemistry and molecular biology [39 (link), 40 (link)]. In this study, Western blot was used in detecting the expression of AEG-1 protein in different cell lines and different groups. The NSCLC cells were washed with PBS in a 6-well-plate and were lysed in a buffer of 10% glycerol, 5% SDS, 5% 2-mercaptoethanol, 0.2% romophenol blue, 80 mM Tris-HCl (ph 6.8), 5 mM EDTA (ph 8) and 1 mM phenylmethylsulfonyl fluoride. Then, the lysates were centrifuged at 12,000×g for 10 min at 4°C and were boiled for 5 min. Hybond ECL nitrocellulose membranes (GE Healthcare Bio-sciences/Amersham, Diegem, Belgium) were applied to separate the proteins for 2 h at 100 mA. The immunoblot analyses were executed using the following antibodies: AEG-1 (Cell Signaling Technology) and GAPDH (Vazyme Biotech Co., Ltd). The image pro plus 6.0 was used to analyze the IOD value.