Product DescriptionMouse anti-Methyl-CpG binding protein 2 (MeCP2) Monoclonal Antibody (Unconjugated), suitable for WB, IHC-Frozen.
Application(s)IHC-Frozen, WB
Application DetailsWestern blotting (1:5,000-1:10,000) and Immunohistochemistry (1:1,000-1:5,000). 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 HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG1
Clone Name4F11
ConjugateUnconjugated
Immunogen DescriptionFull-length recombinant 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 rat cerebellum section with mouse antibody to MeCP2 (1:500, red) and rabbit antibody to GFAP (R-1374-50, 1:5,000, green) by Immunohistochemistry. While MeCP2 is specifically expressed in nuclei of neuron cells, the GFAP antibody stains the network of glial cells. Blue: DAPI nuclear stain. IHC method: Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 1 hour, cut to 45 uM, and free-floating sections were stained with above antibodies. 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: Rat cerebellum stained with mouse anti-MeCP2 (red, 1:500) and chicken anti-calbindin (C-1798-50, green, 1:2,000) by Immunohistochemistry. Blue: DAPI nuclear stain. IHC Method: Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 1 hour, cut to 45 um, and free-floating sections were stained. The MeCP2 antibody specifically labels nuclei of neuronal cells to a variable extent, while calbindin is expressed in the dendrites and perikarya of Purkinje cells in the molecular layer of cerebellum. Right: Western blot analysis with mouse anti-MeCP2 (green, 1:1,000) and chicken anti-GAP43 (red). (2) rat whole brain, (3) nuclear fraction of rat brain, (4) mouse whole brain, (5) nuclear fraction of mouse brain. A strong band at about 75 kDa is observed in rat and slightly lower in mouse nuclear enriched fractions, corresponding to MeCP2 protein. GAP43 protein is detected in all preparations with apparent molecular weight of 43 kDa.
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