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

Conjugates logo Related conjugates and formulations

Unconjugated

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. This information is proprietary to Abcam and/or its suppliers.

  • 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.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    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.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

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

Properties

Applications

The Abpromise guarantee

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).
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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