Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)

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Western blot - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
  • Immunocytochemistry - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
  • Western blot - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
  • Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3564] to MBD1 - BSA and Azide free
  • Suitable for: ICC, IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-MBD1 antibody [EPR3564] - BSA and Azide free
    See all MBD1 primary antibodies

  • Description

    Rabbit monoclonal [EPR3564] to MBD1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, IHC-P, WBmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human MBD1 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, A431, 293T and Jurkat whole cell lysate (ab7899). IHC-P: Human tonsil tissue. ICC: MCF7 cells.

  • General notes

    ab247676 is the carrier-free version of ab108510 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab247676 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab247676 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
ICC Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.

 
WB Use at an assay dependent concentration. Detects a band of approximately 80 kDa (predicted molecular weight: 67 kDa).
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      Transcriptional repressor that binds CpG islands in promoters where the DNA is methylated at position 5 of cytosine within CpG dinucleotides. Binding is abolished by the presence of 7-mG that is produced by DNA damage by methylmethanesulfonate (MMS). Acts as transcriptional repressor and plays a role in gene silencing by recruiting AFT7IP, which in turn recruits factors such as the histone methyltransferase SETDB1. Probably forms a complex with SETDB1 and ATF7IP that represses transcription and couples DNA methylation and histone 'Lys-9' trimethylation. Isoform 1 and isoform 2 can also repress transcription from unmethylated promoters.

    • Tissue specificity

      Widely expressed.

    • Sequence similarities

      Contains 3 CXXC-type zinc fingers.
      Contains 1 MBD (methyl-CpG-binding) domain.

    • Domain

      The methyl-CpG-binding domain (MBD) functions both in binding to methylated DNA and in protein interactions.
      The third CXXC-type zinc finger mediates binding to non-methylated CpG dinucleotides.
      The transcriptional repression domain (TRD) is involved in transcription repression and in protein interactions.

    • Post-translational
      modifications

      Sumoylated with SUMO1 by PIAS1 and PIAS3. Sumoylation affects transcriptional silencing by preventing the interaction with SETDB1. In contrast, sumoylation may increase interaction with AFT7IP.
      Phosphorylated upon DNA damage, probably by ATM or ATR.

    • Cellular localization

      Nucleus speckle. Chromosome. Nuclear, in a punctate pattern. Associated with euchromatic regions of the chromosomes, with pericentromeric regions on chromosome 1 and with telomeric regions from several chromosomes.
    • Information by UniProt

    • Database links

      • Alternative names

        • CXXC 3 antibody
        • CXXC-type zinc finger protein 3 antibody
        • CXXC3 antibody
        • MBD 1 antibody
        • Mbd1 antibody
        • MBD1_HUMAN antibody
        • MECP1 COMPLEX antibody
        • Methyl CpG binding domain protein 1 antibody
        • Methyl CpG binding domain protein 1 isoform PCM1 antibody
        • Methyl CpG binding protein MBD1 antibody
        • Methyl CpG binding protein splice variant 1 antibody
        • Methyl CpG binding protein splice variant 2 antibody
        • Methyl CpG binding protein splice variant 3 antibody
        • Methyl CpG binding protein splice variant 4 antibody
        • Methyl-CpG-binding domain protein 1 antibody
        • Methyl-CpG-binding protein MBD1 antibody
        • PCM 1 antibody
        • PCM1 antibody
        • Protein containing methyl-CpG-binding domain 1 antibody
        • Regulator of fibroblast growth factor 2 (FGF 2) transcription antibody
        • RFT antibody
        • The regulator of fibroblast growth factor 2(FGF 2) transcription antibody
        see all

      Images

      • Western blot - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
        Western blot - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
        All lanes : Anti-MBD1 antibody [EPR3564] (ab108510) at 1/500 dilution

        Lane 1 : Wild-type HAP1 cell lysate
        Lane 2 : MBD1 knockout HAP1 cell lysate
        Lane 3 : A431 cell lysate
        Lane 4 : Jurkat cell lysate

        Lysates/proteins at 40 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 67 kDa
        Observed band size: 70-80 kDa
        why is the actual band size different from the predicted?



        This data was developed using the same antibody clone in a different buffer formulation (ab108510).

        Lanes 1 - 4: Merged signal (red and green). Green - ab108510 observed at 70-80 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

        ab108510 was shown to react with MBD1 in wild-type HAP1 cells in western blot with loss of signal observed in MBD1 knockout sample. Wild-type and MBD1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab108510 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 500 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

      • Immunocytochemistry - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
        Immunocytochemistry - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
        This data was developed using ab108510, the same antibody clone in a different buffer formulation.ab108510, at 1/100 dilution, staining MBD1 in MCF7 cells by Immunofluorescence.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
        This data was developed using ab108510, the same antibody clone in a different buffer formulation.ab108510, at 1/100 dilution, staining MBD1 in paraffin-embedded Human tonsil tissue by Immunohistochemistry. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
      • Western blot - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
        Western blot - Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
        All lanes : Anti-MBD1 antibody [EPR3564] (ab108510) at 1/500 dilution

        Lane 1 : HeLa cell lysate
        Lane 2 : A431 cell lysate
        Lane 3 : 293T (Human embryonic kidney epithelial cell) cell lysate
        Lane 4 : Jurkat cell lysate

        Lysates/proteins at 10 µg per lane.

        Predicted band size: 67 kDa



        This data was developed using ab108510, the same antibody clone in a different buffer formulation.

      • Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)
        Anti-MBD1 antibody [EPR3564] - BSA and Azide free (ab247676)

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (0)

      Publishing research using ab247676? Please let us know so that we can cite the reference in this datasheet.

      ab247676 has not yet been referenced specifically in any publications.

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