Overview

  • Product name

    Anti-Dnmt3a antibody [EPR18455-94]
    See all Dnmt3a primary antibodies
  • Description

    Rabbit monoclonal [EPR18455-94] to Dnmt3a
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: ChIP,Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Dnmt3a aa 600-700. The exact sequence is proprietary.
    Database link: Q9Y6K1

  • Positive control

    • WB: HAP1, HEK-293T, HeLa and NIH/3T3 whole cell lysates; rat brain lysate. IHC-P: Human breast carcinoma and placenta tissue; mouse colon tissue; rat colon tissue.
  • 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

  • Form

    Liquid
  • Storage instructions

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

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18455-94
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab227823 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. Detects a band of approximately 125 kDa (predicted molecular weight: 102 kDa).
IHC-P 1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for ChIP,Flow Cyt,ICC/IF or IP.
  • Target

    • Function

      Required for genome wide de novo methylation and is essential for the establishment of DNA methylation patterns during development. DNA methylation is coordinated with methylation of histones. It modifies DNA in a non-processive manner and also methylates non-CpG sites. May preferentially methylate DNA linker between 2 nucleosomal cores and is inhibited by histone H1. Plays a role in paternal and maternal imprinting. Required for methylation of most imprinted loci in germ cells. Acts as a transcriptional corepressor for ZNF238. Can actively repress transcription through the recruitment of HDAC activity.
    • Tissue specificity

      Highly expressed in fetal tissues, skeletal muscle, heart, peripheral blood mononuclear cells, kidney, and at lower levels in placenta, brain, liver, colon, spleen, small intestine and lung.
    • Sequence similarities

      Belongs to the C5-methyltransferase family.
      Contains 1 ADD domain.
      Contains 1 GATA-type zinc finger.
      Contains 1 PHD-type zinc finger.
      Contains 1 PWWP domain.
    • Domain

      The PWWP domain is essential for targeting to pericentric heterochromatin.
    • Post-translational
      modifications

      Sumoylated; sumoylation disrupts the ability to interact with histone deacetylases (HDAC1 and HDAC2) and repress transcription.
    • Cellular localization

      Nucleus. Cytoplasm. Accumulates in the major satellite repeats at pericentric heterochromatin.
    • Information by UniProt
    • Database links

    • Alternative names

      • DNA (cytosine 5) methyltransferase 3 alpha antibody
      • DNA (cytosine 5) methyltransferase 3A antibody
      • DNA (cytosine-5)-methyltransferase 3A antibody
      • DNA cytosine methyltransferase 3A2 antibody
      • DNA methyltransferase 3 alpha antibody
      • DNA methyltransferase 3a antibody
      • DNA methyltransferase HsaIIIA antibody
      • DNA MTase HsaIIIA antibody
      • DNM3A_HUMAN antibody
      • DNMT 3a antibody
      • DNMT antibody
      • Dnmt3a antibody
      • DNMT3A2 antibody
      • M.HsaIIIA antibody
      • MCMT antibody
      • OTTHUMP00000201149 antibody
      • TBRS antibody
      see all

    Images

    • All lanes : Anti-Dnmt3a antibody [EPR18455-94] (ab227823) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : Dnmt3a knockout HAP1 whole cell lysate
      Lane 3 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
      Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 102 kDa



      Blocking and dilution buffer: 5% NFDM/TBST.

      ab227823 was shown to specifically react with wild-type HAP1 cells as signal was lost in Dnmt3a knockout cells. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab227823 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

      The observed bands ranging from 75~125kDa represent different Dnmt3a isoforms.

    • Anti-Dnmt3a antibody [EPR18455-94] (ab227823) at 1/1000 dilution + Rat brain lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 102 kDa
      Observed band size: 125 kDa
      why is the actual band size different from the predicted?


      Exposure time: 26 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

      The observed bands ranging from 75~125kDa represent different Dnmt3a isoforms.

       

    • Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the rat colon (PMID: 24837369) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

      The section was incubated with ab227823 for 30 mins at room temperature.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the mouse colon (PMID: 24837369) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

      The section was incubated with ab227823 for 30 mins at room temperature.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human placenta (PMID: 22413869) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

      The section was incubated with ab227823 for 30 mins at room temperature.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human breast carcinoma (PMID: 24464625) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

      The section was incubated with ab227823 for 30 mins at room temperature.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    References

    ab227823 has not yet been referenced specifically in any publications.

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