Product DescriptionRabbit anti-Methyl-CpG- binding protein 2 (MeCP2) Polyclonal Antibody (Unconjugated), suitable for WB, IHC-Frozen.
Application(s)IHC-Frozen, WB
Application DetailsWestern blotting (1:1,000-1:5,000) and Immunohistochemistry (1:1,000) Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetMethyl-CpG- binding protein 2 (MeCP2)
SpecificityHuman,reacts with Human, Mouse, Rat.
Target Host SpeciesHuman
Species ReactivityHuman, Mouse, Rat
Antibody HostRabbit
Antibody TypePolyclonal
Antibody IsotypeIgG
ConjugateUnconjugated
Immunogen DescriptionA synthetic peptide (REEPVDSRTPVTERVS, aa: 471-486) of C-terminus of human protein.
Purity DescriptionAffinity purified
FormatLyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, without preservatives
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material. Final buffer contains no preservatives.
Storage InstructionsStore lyophilized antibody at 2-8°C. After reconstitution divide into aliquots and store at -20°C for long-term storage. Store at 2-8°C short-term (up to 4 weeks) with an appropriate antibacterial agent. Avoid repetitive freeze/thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Scientific BackgroundChromosomal protein that binds to methylated DNA. It can bind specifically to a single methyl-CpG pair. It is not influenced by sequences flanking the methyl-CpGs. Mediates transcriptional repression through interaction with histone deacetylase and the corepressor SIN3A. Binds both 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC)-containing DNA, with a preference for 5-methylcytosine (5mC). Ref: uniprot.org
Left: Staining of mouse brain section with rabbit antibody to MeCP2 (green) by Immunohistochemistry. MeCP2 is mainly associated with methylated DNA in nucleus of neuronal cells. Blue: DAPI nuclear stain. IHC method: Following transcardial perfusion of mouse with 4% paraformaldehyde, brain was post fixed overnight and cut to 45 uM. Right: Western blot analysis of MeCP2 expression in nuclear extracts from mouse brain. This antibody recognizes a strong and clear band at 74 kDa corresponding to MeCP2. The molecular weight of MeCP2 is 52 KDa, however, on SDS-PAGE this protein migrates slower due to its unusual high positive charge density.
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). [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 of ~54 kDa.
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