Horseradish peroxidase hrp conjugated goat anti rabbit immunoglobulin g igg antibody sc 2030

Human chondrocytes were plated at 5 ×10⁴ per well into 24 well plates. Cells were treated with CA and/or stimulated with interleukin-1β (IL-1β, 1 ng /ml). After incubation for 24h, the cells were washed with PBS, and proteins were extracted using M-PER™ protein extraction reagent including protease inhibitor cocktail (Thermo Fisher Scientific). Equivalent amounts of protein (15ug/lane) were separated on Mini-PROTEAN TGX gels (Bio-Rad Laboratories, Hercules, CA, USA), and transferred to a polyvinylidene fluoride (PVDF) membrane using Trans-Blot Turbo transfer system (Bio-Rad Laboratories). Anti-HO-1 antibody (1:2000, abcam), anti-GAPDH antibody (1:1000, Millipore) were used as primary antibodies. Horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (IgG) antibody (sc-2030; Santa Cruz Biotechnology, Dallas, TX, USA) and anti-mouse IgG antibody (sc-2005; Santa Cruz Biotechnology) were used as secondary antibody. The signal was detected with chemiluminescence (Wako) using the ImageQuant LAS 4000 system (GE Healthcare).