Anti mouse igg hrp linked antibody

On the 30th day, cortical and striatal brain tissue was extracted from all four groups of mice. The tissue was homogenized in the lysis buffer (150 mM sodium chloride, 1.0 % TritonX-100, 0.5 % sodium dodecyl sulfate, and 50 mM Tris, pH 8.0). The protein concentration was determined using a Bradford protein assay kit (Bio-Rad, USA). We loaded equal amounts of protein (50 μg) run in a 12 % gel, and then transferred to PVDF (Pall Corporation) membrane through a trans blot wet transfer system (Bio-Rad). The membrane was blocked using 5 % BSA and incubated with respective primary and secondary antibodies for α7 nAChRs mouse mAb (CHRNA7, 1:500, #MA5–31691, Thermo Fischer); Anti-mouse IgG-HRP-linked antibody (1:5000, AB_10015289, Jackson ImmunoResearch Laboratories). Membranes stained with ponceau (ML045, Himedia) were used as a control for normalization. The signal intensities of the bands were captured using the fusion pulse gel documentation system (Eppendorf, USA). ImageJ software was used to quantify the band intensities.