Rabbit anti human beta actin antibody

Rabbit anti-human polyclonal EGFR antibody (Cat. number ab2430), rabbit anti-human polyclonal GRB2 antibody (ab32037), rabbit anti-human monoclonal PTPN11 antibody (Cat. number ab32083), and rabbit anti-human beta-actin antibody (Cat. number ab8227) and goat anti-rabbit HRP (IgG H&L) (Cat. number ab6721) were purchased from Abcam Shanghai office launch (Shanghai, China). The intestinal samples were taken from all participants by a noninvasive method using endoscopic techniques. All tissues were ground by using a sterile mortar and pestle. Sample proteins were collected by centrifugation and separated by SDS-PAGE and then electrophoretically transferred onto PVNF membranes. After blocking the membranes with free-fat milk, they were then incubated with primary antibody. The members were washed three times and incubated with HRP-linked secondary antibody. The protein expression level was normalized by beta-actin expression. The immunoreactive result was visualized by using an enhanced chemiluminescence system (Amersham Pharmacia Biotech, Stockholm, Sweden).

Ten milligrams of fetal membranes or placental villi were dissected and lysed in RIPA buffer containing protease inhibitors (Roche, United States). Western blot analysis was performed by loading 15 μg of proteins on 4–12% pre-cast Bis-Tris gels (Bio-Rad, United States) and transferred to PVDF membranes (Merck Millipore, United States). Membranes were incubated with mouse anti-human MPO monoclonal antibody (Santa Cruz, United States, 1:200 diluted in 2% milk), mouse anti-human elastase, neutrophil expressed (ELANE) monoclonal antibody (Santa Cruz, United States, 1:200 diluted in 2% milk), mouse anti-human GAPDH monoclonal antibody (Thermo Fisher Scientific, United Kingdom, 1:2,000 diluted in 2% milk), rabbit anti-human beta-actin antibody (Abcam, United Kingdom, 1:1,000 diluted in 2% milk). Dye-800-conjugated secondary antibodies were applied and visualized with an Odyssey Clx (Li-Cor, United States). Image studio (Li-Cor, United States) and Image J (Schneider et al., 2012 (link)) were used for Western blot quantitation, and one-way ANOVA was used for statistical significance test.