Anti mecp2 antibody

Manufactured by Abcam

Sourced in United Kingdom

The Anti-MeCP2 antibody is a tool used in molecular biology research to detect and analyze the expression of the Methyl-CpG Binding Protein 2 (MeCP2) in various samples. MeCP2 is a transcriptional regulator that binds to methylated DNA and plays a crucial role in gene expression and chromatin remodeling. This antibody can be utilized in techniques such as Western blotting, immunohistochemistry, and immunocytochemistry to measure the presence and distribution of MeCP2 in cells and tissues.

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Lab products found in correlation

Orange chip kit, by Diagenode (1 mentions) Formaldehyde, by Merck Group (1 mentions) Chip it express kit, by Active Motif (1 mentions) Rabbit anti mecp2 antibody, by Cell Signaling Technology (1 mentions) Rabbit anti creb antibody, by Cell Signaling Technology (1 mentions) Protein a g agarose bead, by Santa Cruz Biotechnology (1 mentions)

3 protocols using anti mecp2 antibody

ChIP-qPCR Analysis of H19-DMR

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Chromatin immunoprecipitation (ChIP) experiments were performed by using the Orange ChIP kit (Diagenode, Belgium), according to the manufacturer’s instructions. Briefly, cross-linking between DNA and protein was fixed in homogenized endometrial tissues by adding formaldehyde (37%; Sigma, USA). Next, by adding glycine, the cross-linking reaction induced by formaldehyde was quenched. Sonication was used to fragment chromatin to an average DNA fragment size of 200–600 bp using the Bioruptor Sonication System (Diagenode, UCD 200 Bioruptor). One percent of the sheared chromatin was saved, as control input DNA (without adding any antibody), while the rest was incubated with anti-MeCP2 antibody (Abcam, UK) for immunoprecipitation. Real time quantitative PCR (qPCR) was used to analyze level of DNA methylation modifications of H19-DMR region with specific primer sets (Table 1). The primers were designed to amplify two different regions of H19-DMR. Data is reported based on the fold enrichment of different immunoprecipitated DNA relative to 1/100 dilution of input chromatin. The % input was determined using the following formula: % input = 2^{(CtP inputP – CtP ChIPP)} B B × Fd × 100%.

Kamrani S., Amirchaghmaghi E., Ghaffari F., Shahhoseini M, & Ghaedi K. (2022). Altered gene expression of VEGF, IGFs and H19 lncRNA and epigenetic profile of H19-DMR region in endometrial tissues of women with endometriosis. Reproductive Health, 19, 100.

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Chromatin Immunoprecipitation Assay for MeCP2 and H3K9,14ac

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Bilateral dissections of the mPFC were performed on wet ice and the tissue was cross-linked in 1% formaldehyde. Tissue was lysed with a hand-held homogenizer in cell lysis buffer (100mM EDTA, 50mM Tris-HCl, 1% SDS) containing protease inhibitors and centrifuged at 5000rpm. The nuclear pellet was re-suspended in a 2:1 ratio of ChIP dilution buffer (0.01% SDS, 1.2mM EDTA, 16.7mM Tris-HCl, 167mM NaCl, 1.1% Triton X-100) and nuclear lysis buffer (10mM NaCl, 10mM Tris-HCl, 3mM MgCl2, 1% NP-40) and fragmented by sonication. Samples were centrifuged at maximum speed and the supernatant was collected and stored at -80°C. An aliquot of the fragmented chromatin was set aside as total input control.
ChIP assays were performed using the ChIP-IT Express kit (Active Motif, Carlsbad, CA) following the protocol supplied by the company. For the ChIP reaction, 100ul of the fragmented chromatin was mixed with protein G magnetic beads and 5ul of anti-MeCP2 antibody (Abcam, Cambridge, MA) or anti-acH3K9,14 antibody (Millipore, Billerica, MA). The reaction mixture was incubated at 4°C for 24 hrs on a rotator. The beads were washed with wash buffer and the DNA was immunoprecipitated.

Kabir Z.D., Kennedy B., Katzman A., Lahvis G, & Kosofsky B.E. (2014). Effects of prenatal cocaine exposure on social development in mice. Developmental neuroscience, 36(0), 338-346.

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Immunoprecipitation and Immunoblotting of MeCP2, CREB, G-quadruplex, and hnRNPA1

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Immunoprecipitation and immunoblotting analysis were performed as previously described. Briefly, TM3 cells were lysed with SDS lysis buffer at 4°C for 10 min. For immunoprecipitation, lysates were incubated overnight at 4 °C with anti-MeCP2 antibody (Abcam, Cat# ab2828), rabbit anti-CREB antibody (Cell Signaling, Cat# 9197 S), anti-DNA G-quadruplex structures antibody, clone BG4 (Sigma-Aldrich, Cat# MABE917), and rabbit anti-HnRNPA1(Cell Signaling, Cat# 8443), and then with Protein A + G Agarose beads (Santa Cruz Biotechnology, Cat# sc-2003) at 4°C for 2 h. Subsequently, immunoprecipitants were washed three times, equivalent amounts of protein were detected by Western blot with rabbit anti-MeCP2 antibody (Cell Signaling Technology; Cat# 3456; 1:1000).

Wang Y.M., Wu Y., Zheng Y.F, & Wang H.Y. (2021). MeCP2 duplication causes hyperandrogenism by upregulating LHCGR and downregulating RORα. Cell Death & Disease, 12(11), 999.

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