| Name: | Rabbit polyclonal antibody to MeCP2 |
| Immunogen: | Full length recombinant human MeCP2 |
| HGNC Name: | MECP2 |
| UniProt: | P51608 |
| Molecular Weight: | 70-75kDa by SDS-PAGE |
| Host: | Rabbit |
| Isotype: | |
| Species Cross-Reactivity: | Human, monkey, rat, mouse |
| RRID: | AB_2572345 |
| Format: | Affinity purified at 1mg/mL in PBS, 50% glycerol, 5mM NaN3 |
| Applications: | WB, IF/ICC, IHC |
| Recommended Dilutions: | WB: 1:10,000-1:20,000 IF/ICC or IHC: 1:2,000 |
| Storage: | Stable at 4°C for one year, for longer term store at -20°C |
Immunofluorescent analysis of rat cerebellum section costained with rabbit pAb to MeCP2, RPCA-MeCP2, dilution 1:5,000, in red, and chicken pAb to calbindin, CPCA-Calb, dilution 1:2,000, in green. Blue is DAPI staining of nuclear DNA. Following transcardial perfusion with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45μM, and free-floating sections were stained with above antibodies. The MeCP2 antibody selectively stains nuclei of neuronal cells to a variable degree. Calbindin, often used as Purkinje cell marker, is prominently expressed in dendrites and perikarya of these cells in the cerebellar molecular layer.
Western blot analysis of tissue and cell lysates using rabbit pAb to MeCP2, RPCA-MeCP2, dilution 1:20,000 in green: [1] protein standard (red), [2] Rat whole brain, [3] nuclear fraction of rat brain, [4] mouse whole brain, [5] nuclear fraction of mouse brain, [6] HeLa cells, [7] 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 which is ~54kDa
Rabbit Polyclonal Antibody to MeCP2
Cat# RPCA-MeCP2
$120.00 – $800.00
Methyl-CpG Binding Protein 2 (MeCP2) is widely expressed in tissues and particularly heavily in neurons. It is a nuclear protein which was found to bind methylated cytosines in CpG islands in DNA, but which may also bind hydroxymethyl cytosine residues (1-3). Methyl and hydroxymethyl cytosines are generally found in regions of DNA regulating the expression of specific genes and MeCP2 is thought to function as a gene regulator, though it may have other functions. The MeCP2 protein contains one methyl-CpG binding domain and two AT hook domains, highly basic peptides which bind AT rich DNA sequences. The MeCP2 gene, MECP2 is located on the X-chromosome and loss of function mutations of the gene cause Rett syndrome, a relatively rare neurodevelopmental, autistic disorder which presents almost exclusively in females, since similar mutation of MECP2 in males is almost invariably lethal (3). Rett patients are apparently normal at birth but develop neurological problems over the period when extensive synaptogenesis is occurring, which coincides with the expression of high levels of MeCP2 in normal brain. The Rett brain shows smaller more densely packed neurons with less extensive dendritic branching (2). Duplication of the MECP2 gene was found to be the cause of a different neurodevelopmental disorder referred to a MECP2 duplication syndrome (4). Levels of MeCP2 normally alter dynamically and the protein is regulated by phosphorylation on multiple sites (5-7).
The RPCA-MeCP2 antibody was made against full length recombinant human MeCP2 expressed in and purified from E. coli and can be used to identify neurons transiently expressing high levels of this nuclear protein. We also supply a mouse monoclonal and a chicken polyclonal antibody to the same protein, MCA-4F11 and CPCA-MeCP2. Mouse select image at left for larger view.
Chromogenic immunostaining of a 4% PFA fixed paraffin embedded rat hippocampus section with rabbit pAb to MeCP2, RPCA-MeCP2, dilution 1:5,000, detected with DAB (brown) using the Vector Labs ImmPRESS method and reagents with citra buffer retrieval. The RPCA-c-FOS antibody specifically labels activated neurons. This antibody performs well with 4% PFA and NBF fixed mouse, and rat tissues; but is weak in human sections tested due to both its rapid turnover rate and sensitivity to over fixation. Mouse select image for larger view.
Immunofluorescent analysis of COS-1 cells costained with rabbit pAb to MeCP2, RPCA-MeCP2, dilution 1:5,000, in red, and mouse mAb to ALDHL1, MCA-4A12, dilution 1:1,000, in green. Blue is DAPI staining of nuclear DNA. Following fixation with 3.7% formalin for 5 min, cells were permeabilized with ice cold methanol, blocked and stained with above antibodies. The MeCP2 antibody selectively stains nuclei of these African green monkey derived cells, while ALDH1L1 protein is expressed in the cytoplasm of these cells. Mouse select image for larger view.
1. Klose RJ, et al. DNA binding selectivity of MeCP2 due to a requirement for A/T sequences adjacent to methyl-CpG. Mol. Cell 19:667–78 (2005).
2. Amir RE, et al. Rett syndrome is caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat. Genet. 23:185–8 (1999).
3. Pohodich AE, Zoghbi HY. Rett syndrome: disruption of epigenetic control of postnatal neurological functions. Hum. Mol. Genet. 24:R10-6 (2010).
4. Ramocki MB, Tavyev YJ, Peters SU. The MECP2 duplication syndrome. Am. J. Med. Genet. A. 152A:1079-88 (2010).
5. Zhou Z, et al. Brain-specific phosphorylation of MeCP2 regulates activity dependent Bdnf transcription, dendritic growth, and spine maturation. Neuron 52:255–69 (2006).
6. Deng JV, et al. MeCP2 in the nucleus acumbens contributes to neural and behavioral responses to psychostimulants. Nat. Neurosci. 13:1128-36 (2010).
7. Tao J, et al. Phosphorylation of Mecp2 at Serine 80 regulates its chromatin association and neurological function. PNAS 24:106 (2009).
