Dako animal research kit for mouse primary antibodies

Cerebrum, heart and kidney were formalin-fixed and paraffin-embedded. Multiple sections (4 μm) were deparaffinized with xylene and stained with hematoxylin and eosin (H&E), Masson's trichrome (TCM), periodic acid–Schiff (PAS) and Luxol fast blue (LFB) (Garcia-Corzo et al, 2013 (link)). Immunohistochemistry was carried out in the same sections, using the following primary antibodies: glial fibrillary acidic protein or anti-GFAP (Millipore, MAB360), anti-oligodendrocytes (Millipore, MAB1580) and neuronal class III β-tubulin anti-TUJ1 (Covance, MMS-435P) (Garcia-Corzo et al, 2013 (link)). Dako Animal Research Kit for mouse primary antibodies (Dako Diagnóstico S.A., Spain) was used for the qualitative identification of antigens by light microscopy. Sections were examined at 40–400× magnifications with an OLYMPUS CX41 microscope, and the images were scanned under equal light conditions with the CELL A computer program.
Muscle samples (triceps surae) were snap-frozen in isopentane cooled in liquid nitrogen. Cross sections (8 μm thick) of frozen muscle were stained for succinate dehydrogenase (SDH) and cytochrome oxidase (COX) activities (Tanji & Bonilla, 2008 (link)). Muscle sections were also stained with hematoxylin–eosin and Gomori Trichrome to assess muscle fiber area and general morphology (Tanji & Bonilla, 2008 (link)).

Liver were formalin-fixed and paraffin-embedded. Multiple sections (4 μm) were deparaffinized with xylene. Immunohistochemistry was carried out using anti-8OHdG (QED Bioscience, 12501) as primary antibody^{17 (link)}. Dako Animal Research Kit for mouse primary antibodies (Dako Diagnóstico S.A., Spain) was used for the qualitative identification of antigens by light microscopy. Sections were examined at 10× magnifications with a ZEISS Primo Star microscope, and the images were scanned under equal light conditions with the Axio Vision 4.8.2 computer program. The 8OHdG positive signal was quantified by using the software ImageJ (National Institutes of Health, USA) and the results were expressed by the percentage of the positive signal.

Mice brains were formalin fixed and paraffin embedded. Multiple sections (4 μm thickness) were deparaffinized with xylene and stained with hematoxylin and eosin (H&E). Immunohistochemistry was carried out in the same sections, using the following primary antibodies: Glial fibrillary acidic protein or anti-GFAP (Millipore, MAB360), and anti-Iba-1 (Wako, 019-19741). Dako Animal Research Kit for mouse primary antibodies (Dako Diagnóstico S.A., Spain) was used for the qualitative identification of antigens by light microscopy. Sections were examined at 40–400 magnifications with an OLYMPUS CX41 microscope, and the images were scanned under equal light conditions with the CELL A computer program [4 (link)]. The GFAP positive signal was quantified by using the software ImageJ (National Institutes of Health, USA) and the results were expressed by the percentage of the positive signal.