This is a knockout-validated antibody summary, based on the publication "Brain phosphorylation of MeCP2 at serine 164 is developmentally regulated and globally alters its chromatin association", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Total brain and tissue lysates from WT and MeCP2-KO mice were subjected to western blot and incubated with antibodies to total MeCP2. Tubulin was used as loading control. From [1].

Rabbit polyclonal

Company: Sigma

Antibody: MeCP2

Catalog number: M9317

Summary: Rabbit polyclonal against a synthetic peptide corresponding to the C-terminus of human MeCP2 (amino acids 465-478). Reacts with human by western blot and microarray.

Western blot | Immunocytochemistry

WB: Tissues from WT and MeCP2-KO mice. Samples were homogenized in lysis buffer (50 mM Tris HCl pH 7.4, 150 mM NaCl, 1% Triton X-100, 2 mM EDTA, 1 mM DTT) supplemented with PhosSTOP (Roche), cOmplete EDTA-free protease inhibitor cocktail (Roche) with a dounce homogenizer. ICC: cortical neurons, DIV7 and 14, from WT and MeCP2-KO mice. Cells were fixed with 4% PFA for 20 min at RT.

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Figure 2. Representative images from DIV7 neurons prepared from WT or Mecp2 KO E15 cortices. For each condition, under the GFP panel, the three squares show magnification of nuclei stained respectively with: MeCP2 (left), DAPI (centre); merged image is on the right. KO neurons were infected at 0 DIV with MeCP2 WT (KO+WT) or the S164A (KO+S164A) and S164D (KO+S164D) derivatives. From [1].

WB: TBST (10 mM Tris–HCl, pH 8.0, 150 mM NaCl and 0.05% Tween-20) containing 5% milk.

ICC: 5% Horse serum, 0.2% Triton-X-100 in TBS for 1h.

WB: 1:1,000 dilution overnight at 4 °C.

ICC: 1:200 dilution overnight at 4 °C.

WB: appropriate secondary antibody (Life Technologies) for 1 hour at RT.

ICC:Anti-mouse Alexa Fluor 488 for 1 h at RT.

WB: enhanced chemiluminescence (EuroClone).

ICC: NikonEclipse Ni upright microscope.

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.