MeCP2 Polyclonal Antibody

$Nuclear extracts (40 \u03bcg) from HeLa cells were analysed by Western blot using the Bioss antibody against MeCP2 (Cat. No. bs-53112R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.$

Nuclear extracts (40 \u03bcg) from HeLa cells were analysed by Western blot using the Bioss antibody against MeCP2 (Cat. No. bs-53112R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

$Whole cell extracts (40 \u03bcg) from HeLa cells transfected with MeCP2 siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Bioss antibody against MeCP2 (Cat. No. bs-53112R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.$

Whole cell extracts (40 \u03bcg) from HeLa cells transfected with MeCP2 siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Bioss antibody against MeCP2 (Cat. No. bs-53112R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against MeCP2 (Cat. No. bs-53112R) and the crude serum. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be: 1:32,900.

$ChIP assays were performed using human osteosarcoma (U2OS) cells, MeCP2 Polyclonal Antibody (Cat. No. bs-53112R) and optimized PCR primer sets. Sheared chromatin from 1x10e6 cells and 5 \u03bcg of antibody were used per ChIP experiment. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the ZMYND10 gene (used as a positive control) and CDC6 gene (used as a negative control). Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

ChIP assays were performed using human osteosarcoma (U2OS) cells, MeCP2 Polyclonal Antibody (Cat. No. bs-53112R) and optimized PCR primer sets. Sheared chromatin from 1x10e6 cells and 5 \u03bcg of antibody were used per ChIP experiment. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the ZMYND10 gene (used as a positive control) and CDC6 gene (used as a negative control). Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Applications

WB
ELISA
dot-blot

Reactivity

Human

Overview
Catalog #	bs-53112R
Product Name	MeCP2 Polyclonal Antibody
Applications	WB, ELISA, dot-blot
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Polyclonal antibody raised in rabbit against MeCP2 (Methyl-CpG-binding domain protein 2), using a KLH-conjugated synthetic peptide containing a sequence from the C-terminal part of the protein
Clonality	Polyclonal
Isotype	Not Determined
Concentration	1.2ug/ul
Purification	Affinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.
Storage Buffer	PBS with 0.05% sodium azide and 0.05% Proclin300.
Storage Condition	Store at -20°C for 12 months.
Target
Gene ID	4204
Swiss Prot	P51608
Synonyms	Methyl-CpG-binding protein 2, MECP2, MeCp-2 protein
Background	Chromosomal 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).
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
dot-blot