Goat anti mouse igm secondary antibody hrp conjugate

Nuclear lysate from human PBMC was prepared as previously described by [52 (link)]. Twenty μg of nuclear proteins, quantified with the Bradford method (Bio-Rad Laboratories Inc., Hercules, CA, USA), were separated by gel electrophoresis on 4–12% Bis-Tris Criterion XT precast gels (Bio-Rad Laboratories Inc., Hercules, CA, USA) and electroblotted onto polyvinylidene fluoride membranes (Amersham Pharmacia Biotech Inc., Piscataway, NJ, USA). Immunoblotting was performed with rabbit HIF-1α antibody (1:1000), anti-NRF2 polyclonal antibody raised against a peptide mapping at the C-terminus (C-20) (1:1000), rabbit anti-NF-κB (p65) polyclonal antibody (1:1000) (Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-Lamin B monoclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA) (1:1000), followed by peroxidase-conjugated secondary antibody HRP labeled goat anti-rabbit Ig (BD Pharmigen, San Diego, CA, USA) (1:5000), goat anti-mouse IgM secondary antibody HRP conjugate (Thermo Scientific, Waltham, MA, USA) (1:10,000), and visualized with an Electrochemiluminescence (ECL)western blotting system (AmershamBiosciences, Buckinghamshire, UK).

Nuclear lysate from human PBMCs was prepared as previously described by Fratantonio et al. [33 (link)]. In total, 20 μg of nuclear proteins, quantified with the Bradford method (Bio-Rad Laboratories Inc., Hercules, CA, USA), was separated by gel electrophoresis on 4–12% Bis-Tris Criterion XT precast gels (Bio-Rad Laboratories Inc., Hercules, CA, USA) and electroblotted onto polyvinylidene fluoride membranes (Amersham Pharmacia Biotech Inc., Piscataway, NJ, USA). Immunoblotting was performed with rabbit PGC-1α antibody (1:1000), rabbit Prx3 antibody (1:1000), and mouse anti-Lamin B antibody (1:1000) (Santa Cruz Biotechnology, Dallas, TX, USA), followed by peroxidase-conjugated secondary antibody HRP labeled goat anti-rabbit Ig (BD Pharmigen, San Diego, CA, USA) (1:5000) and goat anti-mouse IgM secondary antibody HRP conjugate (Thermo Scientific, Waltham, MA, USA) (1:10,000), and visualized with an Electrochemiluminescence (ECL) Western blotting system (Amersham Biosciences, Buckinghamshire, UK).