Left: Analysis of rat cerebellum with rabbit anti-MeCP2 (red, 1:5,000) and chicken anti-calbindin (C-1798-50, green, 1:2,000) by Immunohistochemistry. Blue: DAPI nuclear stain. IHC Method: Following transcardial perfusion with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45 um, and free-floating sections were stained. The MeCP2 antibody selectively stains nuclei of neuronal cells. Calbindin, often used as Purkinje cell marker, is prominently expressed in dendrites and perikarya of these cells in the cerebellar molecular layer. Right: Western blot analysis of tissue and cell lysates using rabbit anti-MeCP2 (1:20,000).  rat whole brain,  nuclear fraction of rat brain,  mouse whole brain,  nuclear fraction of mouse brain,  HeLa cells,  C6 cells. Major band at about 70-75 kDa corresponds to MeCP2 protein, predominantly detected in the nuclear fraction of the lysates. The apparent molecular weights of MeCP2 based on SDS-PAGE mobility are variable between species and higher than the actual MeCP2 molecular weight of ~54 kDa.