Anti tubulin t6199

Western blot analyses were conducted as previously described (Gonzalez et al., 2014). Briefly, mPFC were quickly removed and stored at −70°C for western blot analyses. Protein samples (30-50 μg) were separated by 12% SDS-PAGE, and the separated proteins transferred to a PVDF membrane. Blots were incubated with primary antibodies: 1:3000 anti-H3ac 06-599 Millipore, 1:3000 anti-H4ac 06-866 Millipore, 1:3000anti-HDAC1 05-100-I Millipore, 1:1000 anti-HDAC2 sc-7899 (H54) Santa Cruz, 1:1000 anti- NMDAζ1 sc-1467 (C20) Santa Cruz, and 1:6000 anti-tubulin T6199 Sigma. Immune complexes were detected with secondary antibodies and chemiluminescence reagents (Amersham, NJ, USA). Bands were then visualized using an Amersham Imager 600 equipped with automatic detection. The resulting images were quantified with ImageJ (NIH) software.

Platelets were lysed in RIPA buffer with protease and phosphatase inhibitors and protein quantification was determined by bicinchoninic acid (BCA) analysis. Anti-SOD2 antibody (ADI-SOD-111-D) was purchased from Enzo Biochem Inc. (Farmingdale, NY), Anti-Tubulin (T6199) was purchased from (Sigma Aldrich, St. Louis MO). Cytochrome C (11940) was purchased from (Cell signaling technology, Danvers MA). Densitometry was analyzed using ImageJ.