Overview

  • Product name

    Anti-Dnmt1 antibody [EPR3521(2)]
    See all Dnmt1 primary antibodies
  • Description

    Rabbit monoclonal [EPR3521(2)] to Dnmt1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Dnmt1 aa 500-600. The exact sequence is proprietary.
    Database link: P26358

  • Positive control

    • WB: Jurkat, HEK-293T, HeLa, and HuT-78 cell lysates. IHC-P: Human stomach and tonsil tissues. ICC/IF: Wild-type HAP1 cells. Flow Cyt: HeLa cell lysate
  • General notes

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

     

    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.

    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. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3521(2)
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab134148 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/IF 1/250.
WB 1/1000 - 1/10000. Predicted molecular weight: 183 kDa.
IHC-P 1/250 - 1/700. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt 1/20.

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

Target

  • Function

    Methylates CpG residues. Preferentially methylates hemimethylated DNA. Associates with DNA replication sites in S phase maintaining the methylation pattern in the newly synthesized strand, that is essential for epigenetic inheritance. Associates with chromatin during G2 and M phases to maintain DNA methylation independently of replication. It is responsible for maintaining methylation patterns established in development. DNA methylation is coordinated with methylation of histones. Mediates transcriptional repression by direct binding to HDAC2. In association with DNMT3B and via the recruitment of CTCFL/BORIS, involved in activation of BAG1 gene expression by modulating dimethylation of promoter histone H3 at H3K4 and H3K9.
  • Tissue specificity

    Ubiquitous; highly expressed in fetal tissues, heart, kidney, placenta, peripheral blood mononuclear cells, and expressed at lower levels in spleen, lung, brain, small intestine, colon, liver, and skeletal muscle. Isoform 2 is less expressed than isoform 1.
  • Sequence similarities

    Belongs to the C5-methyltransferase family.
    Contains 2 BAH domains.
    Contains 1 CXXC-type zinc finger.
  • Domain

    The N-terminal part is required for homodimerization and acts as a regulatory domain.
  • Post-translational
    modifications

    Sumoylated; sumoylation increases activity.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • ADCADN antibody
    • AIM antibody
    • CXXC finger protein 9 antibody
    • CXXC-type zinc finger protein 9 antibody
    • CXXC9 antibody
    • DNA (cytosine 5 ) methyltransferase 1 antibody
    • DNA (cytosine-5)-methyltransferase 1 antibody
    • DNA methyltransferase 1 antibody
    • DNA methyltransferase HsaI antibody
    • DNA methyltransferase M.HsaI. antibody
    • DNA MTase antibody
    • DNA MTase HsaI antibody
    • DNMT 1 antibody
    • DNMT antibody
    • Dnmt1 antibody
    • DNMT1_HUMAN antibody
    • Dnmt1o antibody
    • FLJ16293 antibody
    • HSN1E antibody
    • M.HsaI antibody
    • MCMT antibody
    • Met1 antibody
    • MGC104992 antibody
    • mMmul antibody
    • MommeD2 antibody
    see all

Images

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: DNMT1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: HEK293 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab134148 observed at 183 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab134148 was shown to recognize DNMT1 when DNMT1 knockout samples were used, along with additional cross-reactive bands. Wild-type and DNMT1 knockout samples were subjected to SDS-PAGE. ab134148 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti-Rabbit and 680CW Goat anti-Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • ab134148 staining Dnmt1 in wild-type HAP1 cells (top panel) and DNMT1 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab134148 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling Dnmt1 with purified ab134148 at 1/700. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Anti-Dnmt1 antibody [EPR3521(2)] (ab134148) at 1/5000 dilution (purified) + Jurkat whole cell lysate at 10 µg

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

    Predicted band size: 183 kDa
    Observed band size: 183 kDa



    Blocking and dilution buffer: 5% NFDM /TBST.

  • Flow cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Dnmt1 (red) with unpurified ab134148 at a 1/20 dilution (10ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Anti-Dnmt1 antibody [EPR3521(2)] (ab134148) at 1/5000 dilution (purified) + 293T whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 183 kDa
    Observed band size: 183 kDa



    Blocking and dilution buffer: 5% NFDM /TBST.

  • All lanes : Anti-Dnmt1 antibody [EPR3521(2)] (ab134148) at 1/1000 dilution (unpurified)

    Lane 1 : Jurkat cell lysates
    Lane 2 : 293T cell lysates
    Lane 3 : HeLa cell lysates
    Lane 4 : HuT-78 cell lysates

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 183 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labelling Dnmt1 with unpurified ab134148 at a dilution of 1/250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling Dnmt1 with unpurified ab134148 at a dilution of 1/250.

References

This product has been referenced in:

  • Zhang W  et al. The correlation between DNMT1 and ERa expression and the methylation status of ERa, and its clinical significance in breast cancer. Oncol Lett 11:1995-2000 (2016). IHC-P ; Human . Read more (PubMed: 26998112) »
See 1 Publication for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (pancreatic cancer xenografts)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization
No
Specification
pancreatic cancer xenografts
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 23°C
Fixative
10% NBF

Abcam user community

Verified customer

Submitted Feb 27 2017

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