Validated using a knockout cell line
Recombinant
RabMAb

Anti-MBD3 antibody [EPR9913] (ab157464)

Overview

  • Product name
    Anti-MBD3 antibody [EPR9913]
    See all MBD3 primary antibodies
  • Description
    Rabbit monoclonal [EPR9913] to MBD3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Synthetic peptide corresponding to residues in Human MBD3 (UniProt O95983).

  • Positive control
    • HeLa, 293T, fetal brain and Y79 lysates; HeLa cells; Permeabilized 293T cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab157464 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
WB 1/1000 - 1/5000. Predicted molecular weight: 33 kDa.
ICC/IF 1/100 - 1/1000.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IP Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    Images

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: ProteinX knockout HAP1 cell lysate (20 µg)
      Lane 3: Wild-type HAP1 cell lysate (20 µg)
      Lane 4: ProteinX knockout HAP1 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab157464 observed at 33 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab157464 was shown to recognize MBD3 when MBD3 knockout samples were used, along with additional cross-reactive bands. Wild-type and MBD3 knockout samples were subjected to SDS-PAGE. ab157464 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling MBD3 with purified ab157464 at 1/1000. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

      Control: PBS only

    • All lanes : Anti-MBD3 antibody [EPR9913] (ab157464) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : 293T cell lysate
      Lane 3 : Fetal brain lysate
      Lane 4 : Y79 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 33 kDa

    • Immunofluorescent analysis of HeLa cells labeling MBD3 with ab157464 at 1/100 dilution.
    • Flow cytometric analysis of permeabilized 293T cells labeling MBD3 with ab157464 at 1/10 dilution (red) compared to a rabbit IgG negative control (green).
    • Immunoprecipitation of MBD3, using WT and MBD3-/- ES cells (however this was not a 100% pure MBD3 null population). The protein was immunoprecipitated using 1000µg nuclear extract, and 1.03µg of Rabbit polyclonal to MBD3 (ab157464). The same MBD3 antibody was used for the western blot. Left blot image taken after a short exposure, right image taken after a longer exposure.

      Lanes 1, 3 and 5: Mbd3+/+, Lanes 2, 4 and 6: Mbd3-/- (not 100% pure Mbd3 null population), Lanes 1 and 2: Input, Lanes 3 and 4: IP with ab91458, Lanes 5 and 6: IP with ab157464.

      Band: 33kDa: MBD3



      All lanes : Anti-MBD3 antibody [EPR9913] (ab157464) at 1.03 µg

      All lanes :

      Observed band size: 33 kDa (why is the actual band size different from the predicted?)

      See Abreview

    References

    This product has been referenced in:
    • Patel D  et al. The histone demethylase LSD1 regulates inner ear progenitor differentiation through interactions with Pax2 and the NuRD repressor complex. PLoS One 13:e0191689 (2018). Read more (PubMed: 29370269) »
    • Ma C  et al. SALL1 functions as a tumor suppressor in breast cancer by regulating cancer cell senescence and metastasis through the NuRD complex. Mol Cancer 17:78 (2018). Read more (PubMed: 29625565) »

    See all 5 Publications for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (B lymphoblast)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    10 µg
    Specification
    B lymphoblast
    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

    Submitted Jan 19 2017

    Application
    ChIP
    Sample
    Mouse Cell lysate - whole cell (Embryonic stem cell culture)
    Negative control
    Cell line 7G9 (Mbd3 knockout). Also IgG controls used for no-antibody controls in ChIP
    Specification
    Embryonic stem cell culture
    Detection step
    Real-time PCR
    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
    Positive control
    Cell line 7E12 (flag-tagged Mbd3 allele). Anti-Flag antibody was also used in ChIP for comparison
    Username

    Dr. Robin Floyd

    Verified customer

    Submitted Sep 07 2015

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Sample
    Mouse Cell (MEL)
    Specification
    MEL
    Permeabilization
    Yes - tween-20
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Oct 29 2014

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 6.0 citric acid
    Sample
    Mouse Tissue sections (spleen)
    Specification
    spleen
    Permeabilization
    Yes - tween-20
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Oct 29 2014

    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Mouse Cell lysate - whole cell (MEL)
    Specification
    MEL
    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

    Submitted Oct 29 2014

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
    Sample
    Cow Cell (Bovine fetal fibroblasts)
    Specification
    Bovine fetal fibroblasts
    Permeabilization
    Yes - 0.5% Triton-X 100 in PBS
    Fixative
    Paraformaldehyde
    Username

    Jose Garcia

    Verified customer

    Submitted May 06 2014

    Application
    Immunoprecipitation
    Immuno-precipitation step
    Protein G
    Sample
    Mouse Cell lysate - nuclear (mouse ES cells)
    Specification
    mouse ES cells
    Total protein in input
    1000 µg
    Username

    Abcam user community

    Verified customer

    Submitted Oct 31 2013

    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Sample
    Mouse Cell lysate - nuclear (mouse ES cells)
    Specification
    mouse ES cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 31 2013

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

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