Hrp conjugated goat anti rabbit igg secondary antibody pi1000

Abcam (Cambridge, UK) provided the following primary antibodies: anti-SIRT1 (ab7343; dil. 1:1000), anti-GLO1 (ab171121; dil. 1:1000), anti-SOD1 (ab16831; dil.1:1000), anti-SOD2 (ab86087; dil.1:1000), anti-CAT (ab16731; dil. 1:2000), and anti-β-actin (cat. ab8227; dil. 1:40000). The anti-argpyrimidine antibody (cat. AGE06B; dil. 1:1000) was provided by Nordic-MUbio (Susteren, The Netherlands). The HRP-conjugated goat anti-rabbit IgG secondary antibody (PI1000; dil. 1:1000) was purchased from Vector Laboratories (Burlingame, CA, USA). The peroxidase-conjugated anti-mouse secondary antibody (A9044; dil. 1:1000) was purchased from Sigma-Aldrich (Milan, Italy).

Liver tissues were homogenized in RIPA buffer plus protease and phosphatase inhibitors (Sigma-Aldrich), as suggested by the manufacturer. Denatured protein samples were analyzed through SDS–PAGE and probed with different primary antibodies and horseradish peroxidase–conjugate secondary antibody and detected by ECL ECL Star Enhanced Chemiluminescent Substrate plus (Euroclone). Abcam (Cambridge, UK) provided the following primary antibodies: anti-GLO II (ab154108,); anti-NRF2 (ab76026). The anti-GLO I (sc-67351); anti-CCS (sc-20141); anti CTR1 (sc-66847); anti-ATP7B (sc-373964); anti-COX17 (sc-100521) was purchased from Santa Cruz Biotechnology, Inc, Dallas, TX, USA) and anti-CTR2 (cat. PA5-22961) from Invitrogen (Carlsbad, CA, USA) and anti-ATOX1 (cat. MBS4154812) from Mybiosource (San Diego, CA, USA). The HRP-conjugated goat anti-rabbit IgG secondary antibody (PI1000) was purchased from Vector Laboratories (Burlingame, CA, USA). The peroxidase-conjugated anti-mouse secondary antibody (A9044) was purchased from Sigma-Aldrich (Milan, Italy). The images of the specific immune complexes were revealed, acquired and analyzed by using Enhanced Chemi-luminescent Substrate Kit (cat. EMP001005, Euroclone, Milan, Italy).