Anti mtco1 antibody

The Thermo Scientific Mitochondria Isolation Kit and Nucleus Isolation Kit were used for cell fraction isolation. The extracts were obtained and proteins were solubilized for immunoblotting. The following antibodies were used: HA.11 Clone 16B12 (Covance, MMS-101P), FLAG M2-Peroxidase (HRP) (sigma, M8592), beta-actin (sigma), Anti-ATP5A1 antibody (abcam, ab14748), MYBBP1A Antibody (santa cruz, sc-133800), Anti-PP2A Antibody (Millipore,05-421), Hsp60 (Pierce, MA3-012), Cdk1 Antibody (santa cruz, sc-53219), MSH2 Antibody (Novus, NB100-56428), Anti-Histone H2B antibody (Abcam, ab18977), GAPDH (14C10) (cell signaling, 3683), FANCD2 (novusbio, NB100-182), AIF Antibody (santa cruz, sc-5586), ATAD3A/B/C (santa cruz, sc-292156), EF-Tu (santa cruz, sc-367739), Anti-ALDH2 (sigma, SAB2501484), mtTFA (santa cruz, sc-23588), Anti-MTCO1 antibody (abcam, ab14705). EZview™ Red Anti-HA Affinity Gel (sigma, E6779-1ML), anti-HA beads (Roche), ANTI-FLAG^® M2 Affinity Gel (Sigma, A2220-1ML), Protein G Agarose (Roche, 11719416001) were used for IP.

In preparation for immunohistochemical staining, the 3 µm paraffin sections were dewaxed/deparaffinized and rehydrated. A heat-mediated antigen retrieval procedure in citrate buffer (pH 6.0) was performed. Blocking of endogenous peroxidase was achieved by incubation with 3% H2O2 (Roth, Karlsruhe, Germany) for 10 min at room temperature. Following the blocking with Roti-Block, the sections were incubated with primary antibodies overnight at 4 °C. The following primary antibodies were used: rabbit polyclonal anti-fatty acid synthase (FAS) antibody (Abcam, Camebridge, UK), mouse monoclonal anti-MTCO1 antibody (Abcam, Camebridge, UK) and rabbit polyclonal anti-PPARαantibody (Abcam, Cambridge, UK). After incubation with peroxidase-labeled goat anti-rabbit IgG antibody (KPL, Gaithersburg, MD, USA) or anti-mouse IgG antibody (SeraCare, Milford, MA, USA), di-aminobenzidine (DAB) (DAB-peroxidase substrate kit; Vector Laboratories, Burlingame, CA, USA) was used as a chromogen.
For Oil Red O staining, snap-frozen renal sections (thickness 10 µm) were stained by using Oil Red O solution 0.5% in iso-propanol (Sigma-Aldrich, Merck, Darmstadt, Germany).