Anti bcl xl rabbit monoclonal antibody

Cells were collected in a conical tube and lysed in Laemmli buffer, followed by sonication and heat denaturation. Protein lysates were separated by SDS-PAGE and transferred to polyvinylidene fluoride membranes. Protein-bound membranes were then incubated with blocking buffer containing 5% skim milk in Tris-buffered saline, followed by incubation with following antibodies: anti-RTA mouse monoclonal antibody α50A [13 (link)], anti-ORF57 mouse monoclonal antibody (kindly provided by Drs. Vladimir Majerciak and Zhi-Ming Zheng of National Cancer Institute, MD) [14 (link)–16 (link)], anti-c-Myc rabbit monoclonal antibody (ab32072, Abcam, Cambridge, UK), anti-Bcl-2 rabbit monoclonal antibody (Abcam), anti-Bcl-XL rabbit monoclonal antibody (Abcam), or anti-β-actin (ACTB) mouse monoclonal antibody (AC-15) (Santa Cruz Biotechnology, Inc. Dallas, TX). Membranes were subsequently incubated with anti-mouse IgG goat polyclonal antibody with HRP (Abcam) or anti-rabbit IgG goat polyclonal antibody with HRP (GE Health care, Chicago, Il). Chemiluminescent detection was conducted using a Super Signal West Pico and Femto (Thermo Fisher Scientific, Waltham, MA, USA) and a ChemiDoc imaging system (Bio-Rad, Hercules, CA, USA). Uncropped blot images are shown in Supplementary Fig. S1.

For immunochemistry analysis, hepatic tissue section slides were incubated with anti-BAX rabbit polyclonal antibody (Servicebio biotechnology corp., Wuhan, China) and anti-Bcl-xL rabbit monoclonal antibody (Abcam, Cambridge, UK). After incubation with the primary antibody overnight at 4 °C, the slides were washed with PBS (pH 7.4) 3 times and incubated with goat anti-rabbit secondary antibody for 50 min under room temperature. Then, the slides were stained with diaminobenzidine tetrahydrochloride and counterstained with hematoxylin for 3 min. After dehydration and mounting, the slides were visualized under a Leica microscope (Leica DMi8; Leica Corp., Weztlar, Germany). The image pro plus 6.0 software (Media Cybernetics, Rockville, MD, USA) was used to analyze the Integrated Optical Density (IOD) and area of the positive cells to determine the relative protein expression ratio (IOD/Area).