Left: Analysis of calbindin expression (green, 1:2,000) in rat cerebellum by Immunohistochemistry. Sections were co-stained with rabbit anti-MeCP2 (R-1810-100, red, 1:5,000). IHC Method: Transcardial perfusion with 4% PFA, post fixation for 24 hours. Sections were cut to 45 um and stained (free-floating). Calbindin, often used as a Purkinje cell marker, is prominently detected in dendrites and perikarya of these cells in the cerebellar molecular layer. The MeCP2 antibody selectively stains nuclei of neuronal cells. Right: Western blot analysis of rat cerebellum (2), pig hippocampus (3), cow cerebellum (4), parvalbumin (6), calretinin (7), His-tagged calbindin (8). Chicken anti-calbindin (green, 1:5,000) antibody detects calbindin at ~25 kDa in tissue homogenates (2-4). Recombinant human calbindin protein (8) appears at slightly higher molecular weight due to presence of His-tag. The chicken antibody does not cross-react with closely related protein parvalbumin and calretinin.