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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-MBD2 antibody [EPR18361] (ab188474)

Submit a review Submit a question References (3)
Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Immunocytochemistry/ Immunofluorescence - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Flow Cytometry (Intracellular) - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Immunoprecipitation - Anti-MBD2 antibody [EPR18361] (ab188474)
  • Anti-MBD2 antibody [EPR18361] (ab188474)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18361] to MBD2
  • Suitable for: Flow Cyt (Intra), IHC-P, ICC/IF, IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

APC Carrier Free HRP PE

Overview

  • Product name

    Anti-MBD2 antibody [EPR18361]
    See all MBD2 primary antibodies

  • Description

    Rabbit monoclonal [EPR18361] to MBD2

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HeLa, NIH/3T3, MCF7, A-375 and PC-12 cell lysates; mouse brain, mouse heart and rat brain lysates. IHC-P: Human colon, human gastric cancer, mouse stomach and rat colon tissues. ICC/IF: HepG2 cells. IP: HeLa whole cell lysate.

  • General notes

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab188474 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
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/500.
IP
1/50.
WB
1/1000. Detects a band of approximately 43, 29 kDa (predicted molecular weight: 43 kDa).
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/500.
IP
1/50.
WB
1/1000. Detects a band of approximately 43, 29 kDa (predicted molecular weight: 43 kDa).

Target

  • Function

    Binds CpG islands in promoters where the DNA is methylated at position 5 of cytosine within CpG dinucleotides. Binds hemimethylated DNA as well. Recruits histone deacetylases and DNA methyltransferases. Acts as transcriptional repressor and plays a role in gene silencing. Functions as a scaffold protein, targeting GATAD2A and GATAD2B to chromatin to promote repression. May enhance the activation of some unmethylated cAMP-responsive promoters.

  • Tissue specificity

    Highly expressed in brain, heart, kidney, stomach, testis and placenta.

  • Sequence similarities

    Contains 1 MBD (methyl-CpG-binding) domain.

  • Cellular localization

    Nucleus. Nuclear, in discrete foci. Detected at replication foci in late S phase.
  • Information by UniProt

  • Database links

    • Alternative names

      • Demethylase antibody
      • DMTase antibody
      • MBD 2 antibody
      • Mbd2 antibody
      • MBD2_HUMAN antibody
      • MBD2a antibody
      • Methyl CpG binding domain protein 2 antibody
      • Methyl CpG binding protein MBD2 antibody
      • Methyl-CpG-binding domain protein 2 antibody
      • Methyl-CpG-binding protein MBD2 antibody
      • NY CO 41 antibody
      see all

    Images

    • Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
      Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: MBD2 knockout HAP1 cell lysate (20 µg)
      Lane 3: A375 cell lysate (20 µg)
      Lane 4: Human brain tissue lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab188474 observed at 32 & 49 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab188474 was shown to specifically react with MBD2 when MBD2 knockout samples were used. Wild-type and MBD2 knockout samples were subjected to SDS-PAGE. ab188474 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 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-MBD2 antibody [EPR18361] (ab188474)
      Immunocytochemistry/ Immunofluorescence - Anti-MBD2 antibody [EPR18361] (ab188474)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling MBD2 with ab188474 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HepG2 cell line. The nuclear counter stain is DAPI (blue).

      Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse (AlexaFluor®594) (ab150120) secondary antibody at 1/1000 dilution (red).

      The negative controls are as follows:
      -ve control 1: ab188474 at 1/250 dilution, followed by Goat Anti-Mouse (AlexaFluor®594) (ab150120) secondary antibody at 1/1000 dilution.
      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)

      Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat colon tissue is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Flow Cytometry (Intracellular) - Anti-MBD2 antibody [EPR18361] (ab188474)
      Flow Cytometry (Intracellular) - Anti-MBD2 antibody [EPR18361] (ab188474)

      Intracellular Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling MBD2 with purified ab188474 at 1/70 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

       

       

    • Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
      Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
      All lanes : Anti-MBD2 antibody [EPR18361] (ab188474) at 1/10000 dilution

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate
      Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) lysate
      Lane 3 : MCF7 (Human breast adenocarcinoma cell line) lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Predicted band size: 43 kDa
      Observed band size: 29,43 kDa why is the actual band size different from the predicted?


      Exposure time: 15 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The observed MW is consistent with what has been described in the literature (PMID: 17353267).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)

      Immunohistochemical analysis of paraffin-embedded human colon tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human colon tissue tissue is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
      Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
      All lanes : Anti-MBD2 antibody [EPR18361] (ab188474) at 1/10000 dilution

      Lane 1 : A-375 (Human malignant melanoma cell line) lysate
      Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Predicted band size: 43 kDa
      Observed band size: 29,43 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      5% NFDM/TBST: Blocking and dilution buffer.

      The observed MW is consistent with what has been described in the literature (PMID:17353267).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)

      Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human gastric cancer tissue is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
      Western blot - Anti-MBD2 antibody [EPR18361] (ab188474)
      All lanes : Anti-MBD2 antibody [EPR18361] (ab188474) at 1/1000 dilution

      Lane 1 : Mouse brain lysate
      Lane 2 : Mouse heart lysate
      Lane 3 : Rat brain lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Predicted band size: 43 kDa
      Observed band size: 29,43 kDa why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The observed MW is consistent with what has been described in the literature (PMID:17353267).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD2 antibody [EPR18361] (ab188474)

      Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mouse stomach tissue is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-MBD2 antibody [EPR18361] (ab188474)
      Immunoprecipitation - Anti-MBD2 antibody [EPR18361] (ab188474)

      MBD2 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab188474 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab188474 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
      Lane 1: HeLa whole cell lysate 10µg (Input).
      Lane 2: ab188474 IP in HeLa whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab188474 in HeLa whole cell lysate.
      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 3 seconds.

    • Anti-MBD2 antibody [EPR18361] (ab188474)
      Anti-MBD2 antibody [EPR18361] (ab188474)

    References (3)

    Publishing research using ab188474? Please let us know so that we can cite the reference in this datasheet.

    ab188474 has been referenced in 3 publications.

    • Yue T  et al. MBD2 acts as a repressor to maintain the homeostasis of the Th1 program in type 1 diabetes by regulating the STAT1-IFN-? axis. Cell Death Differ N/A:N/A (2021). PubMed: 34420035
    • Ge Y  et al. Mbd2 Mediates Retinal Cell Apoptosis by Targeting the lncRNA Mbd2-AL1/miR-188-3p/Traf3 Axis in Ischemia/Reperfusion Injury. Mol Ther Nucleic Acids 19:1250-1265 (2020). PubMed: 32074940
    • Gong W  et al. Expression and clinical significance of methyl-CpG binding domain protein 2 in high-grade serous ovarian cancer. Oncol Lett 20:2749-2756 (2020). PubMed: 32782591

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