Anti-MBD2 + MBD3抗体[106B691] - ChIP Grade (ab45027)

概述

  • 产品名称Anti-MBD2 + MBD3抗体[106B691] - ChIP Grade
    参阅全部 MBD2 + MBD3 一抗
  • 描述
    小鼠单克隆抗体[106B691] to MBD2 + MBD3 - ChIP Grade
  • 经测试应用适用于: ICC/IF, IHC-P, WB, Flow Cyt, ChIPmore details
  • 种属反应性
    与反应: Human
    预测可用于: Mouse, Rat, Chicken, Xenopus laevis, Zebrafish
  • 免疫原

    Synthetic peptide:

    CKAFMVTDEDIRKQEE

    , corresponding to amino acids 215-230 of Human MBD3

  • 阳性对照
    • HeLa cells (nuclear fraction).
  • 常规说明

    The theoretical molecular weight of human MBD2 (411 amino acids) is 45 kD and human MBD3 (291 amino acids) is 33 kD.

性能

应用

Our Abpromise guarantee covers the use of ab45027 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/400.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 2 µg/ml. Predicted molecular weight: 33, 45 kDa.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ChIP Use at an assay dependent concentration.

靶标

  • 相关性DNA methylation, or the addition of methyl groups to cytosine bases in the dinucleotide CpG, is imperative to proper development and regulates gene expression. The methylation pattern involves the enzymatic processes of methylation and demethylation. The demethylation enzyme was recently found to be a mammalian protein, which exhibits demethylase activity associated to a methyl-CpG-binding domain (MBD). The enzyme is able to revert methylated cytosine bases to cytosines within the particular dinucleotide sequence mdCpdG by catalyzing the cleaving of the methyl group as methanol. MeCP2 and MBD1 (PCM1) are first found to repress transcription by binding specifically to methylated DNA. MBD2 and MBD4 (also known as MED1) were later found to colocalize with foci of heavily methylated satellite DNA and believed to mediate the biological functions of the methylation signal. Surprisingly, MBD3 does not bind methylated DNA both in vivo and in vitro. MBD1, MBD2, MBD3, and MBD4 are found to be expressed in somatic tissues, but the expression of MBD1 and MBD2 is reduced or absent in embryonic stem cells, which are known to be deficient in MeCP1 activity. MBD4 have homology to bacterial base excision repair DNA N-glycosylases/lyases. In some microsatellite unstable tumors MBD4 is mutated at an exonic polynucleotide tract.
  • 细胞定位Nuclear
  • 数据库链接
  • 别名
    • Demethylase antibody
    • DMTase antibody
    • MBD 2 antibody
    • MBD 3 antibody
    • Methyl CpG binding domain protein 2 antibody
    • Methyl CpG binding domain protein 3 antibody
    • Methyl CpG binding protein MBD2 antibody
    • Methyl CpG binding protein MBD3 antibody
    see all

Anti-MBD2 + MBD3 antibody [106B691] - ChIP Grade 图像

  • All lanes : Anti-MBD2 + MBD3 antibody [106B691] - ChIP Grade (ab45027) at 2 µg/ml

    Lane 1 : HeLa cell lysate (nuclear fraction) without immunising peptide
    Lane 2 : HeLa cell lysate (nuclear fraction) with immunising peptide


    Predicted band size : 33, 45 kDa
    Observed band size : 33,45 kDa
    Additional bands at : 160 kDa,40 kDa. We are unsure as to the identity of these extra bands.
  • ab45027 at 2µg/ml staining MBD2 + MBD3 in human glioma tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed and samples were blocked using 10% BSA. The secondary used was an HRP conjugated anti rabbit/mouse mix.
  • Overlay histogram showing HeLa cells stained with ab45027 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45027, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-MBD2 + MBD3 antibody [106B691] - ChIP Grade (ab45027)参考文献

This product has been referenced in:
  • Teperek M  et al. Sperm and spermatids contain different proteins and bind distinct egg factors. Int J Mol Sci 15:16719-40 (2014). WB ; Frog . Read more (PubMed: 25244019) »
  • Kim JY  et al. Specificity in circadian clock feedback from targeted reconstitution of the NuRD corepressor. Mol Cell 56:738-48 (2014). Read more (PubMed: 25453762) »

See all 6 Publications for this product

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Human glioma cell line D54)
Specification Human glioma cell line D54
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% Triton
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jul 15 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Human glioma)
Specification Human glioma
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 0.01 M Tris HCL, pH 10
Permeabilization No
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Dec 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (Glioma cells)
Specification Glioma cells
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Detection step Semiquantitative PCR
Negative control IgG
Username

Abcam user community

Verified customer

提交于 Jun 15 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (glioma cell line)
Loading amount 20 µg
Specification glioma cell line
Gel Running Conditions Non-reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 13 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"