Horseradish peroxidase hrp conjugated goat anti rabbit igg sc 2004

Specimens, AGS or MKN-45 cells were lysed by radioimmunoprecipitation assay (RIPA) lysis buffer (Beotime, China). After determination, the concentrations of protein samples were adjusted to be equal. Then the protein samples were electrophoresed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by transferred to nitrocellulose membranes (Millipore, USA). Next, the 5% skimmed milk was used to incubate the membranes for 1 h, and then they were incubated with the primary antibodies for EDIL3 (ab88667), E-cadherin (ab40772), N-cadherin (ab76011), Vimentin(ab92547), Snail (ab216347) and GAPDH (ab8245) were purchased from Abcam (Cambridge, MA, USA) for overnight at 4 °C. After washed with tris-buffered saline and Tween 20 (TBST) for 3 times, the membranes were then incubated with horseradish-peroxidase (HRP)-conjugated goat anti-rabbit-IgG (sc-2004, Santa Cruz Biotechnology, Inc.) for 2 h. The bands were assessed by a commercial electrochemiluminescence detection regent (Pierce Biotechnology, USA). Protein levels were determined by normalization to GAPDH.