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Anti-MBD2 + MBD3 antibody [106B691] (ab45027)

Reviews (1)Q&A (1)References (10)
Western blot - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
  • Flow Cytometry (Intracellular) - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)

Key features and details

  • Mouse monoclonal [106B691] to MBD2 + MBD3
  • Suitable for: IHC-P, Flow Cyt (Intra), ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-MBD2 + MBD3 antibody [106B691]

  • Description

    Mouse monoclonal [106B691] to MBD2 + MBD3

  • Host species

    Mouse

  • Tested applications

    Suitable for: IHC-P, Flow Cyt (Intra), ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken, Xenopus laevis, Zebrafish

  • Immunogen

    Synthetic peptide corresponding to Human MBD2 + MBD3 aa 200-300. Synthetic peptide:

    CKAFMVTDEDIRKQEE

    , corresponding to amino acids 215-230 of Human MBD3
    Database link: Q9UBB5
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • HeLa cells (nuclear fraction). IHC-P: Human placenta tissue Flow Cyt (Intra): HeLa cells.

  • General notes

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

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.4
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...

  • Purity

    Protein G purified

  • Primary antibody notes

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

  • Clonality

    Monoclonal

  • Clone number

    106B691

  • Isotype

    IgG1

  • Light chain type

    kappa

  • Research areas

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
IHC-P
1/300.
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF
Use at an assay dependent concentration.
WB (1)
Use a concentration of 2 µg/ml. Predicted molecular weight: 33, 45 kDa.
Notes
IHC-P
1/300.
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF
Use at an assay dependent concentration.
WB
Use a concentration of 2 µg/ml. Predicted molecular weight: 33, 45 kDa.

Target

  • Relevance

    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.

  • Cellular localization

    Nuclear

  • Database links

    see all

  • Alternative names

    • 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

Images

  • Western blot - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
    Western blot - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
    All lanes : Anti-MBD2 + MBD3 antibody [106B691] (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 why is the actual band size different from the predicted?
    Additional bands at: 160 kDa, 40 kDa. We are unsure as to the identity of these extra bands.

  • Flow Cytometry (Intracellular) - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
    Flow Cytometry (Intracellular) - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)Image courtesy of an anonymous Abreview.
    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.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 + MBD3 antibody [106B691] (ab45027)
     
    Immunohistochemistry analysis of immersion fixed paraffin-embedded sections of human placenta labeling MBD2 + MBD3 with  ab45027 at 1/300 for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody. Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue).

     

References (10)

Publishing research using ab45027? Please let us know so that we can cite the reference in this datasheet.

ab45027 has been referenced in 10 publications.

  • Kraushaar DC  et al. The gene repressor complex NuRD interacts with the histone variant H3.3 at promoters of active genes. Genome Res 28:1646-1655 (2018). PubMed: 30254051
  • 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 . PubMed: 26910839
  • Gómez-Del Arco P  et al. The Chromatin Remodeling Complex Chd4/NuRD Controls Striated Muscle Identity and Metabolic Homeostasis. Cell Metab 23:881-92 (2016). PubMed: 27166947
  • 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 . PubMed: 25244019
  • Kim JY  et al. Specificity in circadian clock feedback from targeted reconstitution of the NuRD corepressor. Mol Cell 56:738-48 (2014). PubMed: 25453762
  • Zhu D  et al. Overexpression of MBD2 in glioblastoma maintains epigenetic silencing and inhibits the antiangiogenic function of the tumor suppressor gene BAI1. Cancer Res 71:5859-70 (2011). PubMed: 21724586
  • Mastroeni D  et al. Epigenetic changes in Alzheimer's disease: decrements in DNA methylation. Neurobiol Aging 31:2025-37 (2010). IHC-Fr . PubMed: 19117641
  • Mastroeni D  et al. Epigenetic differences in cortical neurons from a pair of monozygotic twins discordant for Alzheimer's disease. PLoS One 4:e6617 (2009). IHC-FoFr ; Human . PubMed: 19672297
  • Thatcher KN  et al. Homologous pairing of 15q11-13 imprinted domains in brain is developmentally regulated but deficient in Rett and autism samples. Hum Mol Genet 14:785-97 (2005). PubMed: 15689352
  • Balmer D  et al. MECP2 mutations in Rett syndrome adversely affect lymphocyte growth, but do not affect imprinted gene expression in blood or brain. Hum Genet 110:545-52 (2002). PubMed: 12107440

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Western blot abreview for Anti-MBD2 + MBD3 antibody [106B691] - ChIP Grade

 Excellent
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 13 2010

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