This is a knockout-validated antibody summary, based on the publication "Disruption of MeCP2 attenuates circadian rhythm in CRISPR/Cas9-based Rett syndrome model mouse" [1] (discussed below) and article "CREB Signaling Is Involved in Rett Syndrome Pathogenesis" [2] (see figure 1e in the article) and "Losing Dnmt3a dependent methylation in inhibitory neurons impairs neural function by a mechanism impacting Rett syndrome" for western blot knockout validation (see [3] figure 1s1a). Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. (A) Immunohistochemical analysis of MeCP2 in the suprachiasmatic nucleus of wild-type control and Mecp2-knockout (KO) mice. The white rectangle areas in upper panels are magnified in lower panels. (B) Immunoblotting analysis of MeCP2 proteins extracted from differentiated mouse embryonic stem cells. Data of wild-type control and three different cell lines for Mecp2-KO are shown. α-tubulin serves as a loading control. Courtesy of the Authors.

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: MeCP2

Catalog number: 3456

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to the carboxy terminus of human MeCP2. Reacts with human, mouse, rat, and monkey. Suitable for western blot, immunoprecipitation, immunohistochemistry (paraffin) and immunofluorescence/immunocytochemistry.

Western blot | Immunohistochemistry

WB: control and Mecp2-deficient ESCs. Cells were homogenized in an ice-cold RIPA buffer (Wako) supplemented with a protease inhibitor cocktail (Roche) and 1 mM phenylmethylsulfonyl fluoride. IHC: hypothalamus (suprachiasmatic nucleus) of wild-type and a Mecp2 mutant mice. Brains were post-fixed overnight with 4% PFA and 30-μm coronal cryosections were taken (CM1860, Leica Biosystems).

IHC: biotinylated secondary antibodies (BA-1300, Vector Labs).

IHC: avidin–biotin complex system (VECTASTAIN Elite ABC kit, Vector Labs) and DAB kit (Vector Labs).