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

製品の概要

  • 製品名
    Anti-MBD2 + MBD3 antibody [106B691] - ChIP Grade
    MBD2 + MBD3 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [106B691] to MBD2 + MBD3 - ChIP Grade
  • アプリケーション
    適用あり: ICC/IF, IHC-P, WB, Flow Cyt, ChIPmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat, Chicken, Xenopus laevis, Zebrafish
  • 免疫原

    Synthetic peptide:

    CKAFMVTDEDIRKQEE

    , corresponding to amino acids 215-230 of Human MBD3

  • ポジティブ・コントロール
    • HeLa cells (nuclear fraction).
  • 特記事項

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

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 関連性
    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.
  • 細胞内局在
    Nuclear
  • 参照データベース
  • 別名
    • 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 画像

  • 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) 使用論文

This product has been referenced in:
  • Xu M  et al. MeCP2 suppresses LIN28A expression via binding to its methylated-CpG islands in pancreatic cancer cells. Oncotarget 7:14476-85 (2016). WB . Read more (PubMed: 26910839) »
  • 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) »

See all 8 Publications for this product

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Thanks for your call today and for your questions about ab45027. Please see below for information about our testing discount program, and let me know if you have any questions. Since ab45027 has not been tested in rat, I can offer a discount off a futu...

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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
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Abcam user community

Verified customer

投稿 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
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Abcam user community

Verified customer

投稿 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
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Abcam user community

Verified customer

投稿 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

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Verified customer

投稿 May 13 2010

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

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