Horseradish peroxidase conjugated goat anti rabbit g 21234

Antibodies directed against Gα12 (sc-409, sc-515445), Gα13 (sc-293424), ALOX12 (sc-365194), IRE1α (sc-390960), GPX4 (sc-166570), caspase-1 (sc-56036), and F4/80 (sc-52664) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-alpha smooth muscle actin (ab7817) antibody was supplied from Abcam (Cambridge, UK), whereas anti-phospho PERK (Thr980, 3179), anti-PERK (3192), anti-ATF6 (65880), anti-RIP (3493), anti-RIP3 (D4G2A, 95702), cleaved caspase-3 (Asp175, #9664), and IL-1β (3A6, #12242) antibodies were from Cell Signaling Technology (Danvers, MA). Anti-phospho-PERK (Thr980, BS-3330R) was obtained from Bioss (Massachusetts, USA). Horseradish peroxidase-conjugated goat anti-rabbit (G-21234) and goat anti-mouse (G-21040) IgGs were provided from Invitrogen (Carlsbad, CA). DSS (160110) was obtained from MP Biomedicals (formerly ICN Biomedicals), whereas the AZ2 compound was supplied from A Pharma Co. (Goyang, South Korea). Anti-β-actin antibody (A5441) was purchased from Sigma-Aldrich (St. Louis, MO).

The precipitated immune complexes were analyzed by western blotting. After SDS-PAGE, proteins were transferred to the PVDF membrane, which was blocked for 1 h with 5% (w/v) nonfat powdered milk in phosphate-buffered saline (PBS) with 0.2% Tween and stained with anti-binase antibodies [23 (link)] at 1 : 500 at 4°C overnight. EGFR and RAS proteins in cell lysates were detected using anti-pan Ras (1 : 100) (sc-166691; Santa Cruz, USA) and anti-EGFR (1 : 200) antibodies (sc-120; Santa Cruz, USA). The primary antibodies were visualized using horseradish peroxidase-conjugated goat anti-rabbit (G-21234; Invitrogen,Carlsbad, CA, USA) at 1 : 10000 and anti-mouse IgG (H + L) antibodies (ab205719; Invitrogen, Carlsbad, CA, USA) at 1 : 5000 for 1 h at room temperature. Proteins were visualized using an ECL chemiluminescent substrate (Pierce, USA) on a ChemiDocXRS+ gel documenter (Bio-Rad, USA).

PAI-1 (614024) and fibronectin (610077) antibodies were obtained from BD Biosciences (Franklin Lakes, NJ, USA). Phospho-Smad3 (#9520) and Smad2/3 (#3102) antibodies were provided by Cell Signaling (Danvers, MA, USA). CTGF antibodies (SC-101586) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA), and TGF-β1 antibody (NBP1-80289) was obtained from Novus Biologicals (Littleton, CO). Horseradish peroxidase-conjugated goat anti-rabbit (G21234) and anti-mouse (G21040) antibodies were purchased from Invitrogen (Carlsbad, CA, USA). α-SMA (A2547) and β-actin (A5441) antibodies, MTT (M2128), and dimethylsulfoxide (D8418) were acquired from Sigma Chemicals (St. Louis, MO, USA). TGF-β (240-B) was purchased from R&D Systems (Minneapolis, MN, USA). CTGF antibody (sc-101586) obtained from Santa Cruz Biotechnology (Santa Cruz, CA). CCl₄ (33650-0330) was received from Junsei Chemical (Kyoto, CO).