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Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537)

Overview

  • Product nameAnti-Dnmt1 antibody [60B1220.1] - ChIP GradeSee all Dnmt1 primary antibodies ...
  • Description
    Mouse monoclonal [60B1220.1] to Dnmt1 - ChIP Grade
  • SpecificityThis antibody detects a ~180 kDa protein, corresponding to the apparent molecular mass of Dnmt1 on SDS-PAGE immunoblots in samples of human and mouse origin. Immunogen itself has been shown to be toxic.
  • Tested applicationsIHC-P, IHC-Fr, ChIP, Flow Cyt, WB, IP more details
  • Species reactivity
    Reacts with: Mouse, Human, Zebrafish
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide: EKDDREDKENAFKR, corresponding to amino acids 637-650 of Human Dnmt1

  • Positive controlHuman kidney (IHC), mouse ES or NIH 3T3 cell lysate (WB)
  • General notesFor maximum product recovery, centrifuge the product vial before removing cap. Shipped at on gel packs.

Properties

Applications

Our Abpromise guarantee covers the use of ab13537 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
IHC-P IHC-P: Use a concentration of 1 - 2 µg/ml. DNMT1 high levels are toxic, as a result it may be tough to find sometimes. Signal amplification might be needed.
IHC-Fr IHC-Fr: Use at an assay dependent dilution.
ChIP ChIP: Use at an assay dependent dilution.
Flow Cyt Flow Cyt: 1/100.
WB WB: Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 183 kDa).
IP IP: Use 2µg for 106 cells.

Target

  • FunctionMethylates 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 specificityUbiquitous; 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 similaritiesBelongs to the C5-methyltransferase family.
    Contains 2 BAH domains.
    Contains 1 CXXC-type zinc finger.
  • DomainThe N-terminal part is required for homodimerization and acts as a regulatory domain.
  • Post-translational
    modifications
    Sumoylated; sumoylation increases activity.
  • Cellular localizationNucleus.
  • Target information above from: UniProt accession P26358 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
      AIM antibodyCXXC finger protein 9 antibodyCXXC-type zinc finger protein 9 antibody
      CXXC9 antibodyDNA (cytosine 5 ) methyltransferase 1 antibodyDNA (cytosine-5)-methyltransferase 1 antibodyDNA methyltransferase 1 antibodyDNA methyltransferase HsaI antibodyDNA methyltransferase M.HsaI. antibodyDNA MTase antibodyDNA MTase HsaI antibodyDNMT 1 antibodyDNMT antibodyDnmt1 antibodyDnmt1 antibodyDNMT1_HUMAN antibodyDnmt1o antibodyFLJ16293 antibodyHSN1E antibodyM.HsaI antibodyMCMT antibodyMet1 antibodyMGC104992 antibodymMmul antibodyMommeD2 antibody
    see all

Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade images

  • ab13537 at 1/10000 dilution staining Dnmt1 in mouse brain tissue section by Immunohistochemistry (Bouin's fixative fixed paraffin-embedded tissue sections). Tissue underwent heat mediated antigen retrieval in microwave with two, 5 minutes incubation intervals in citrate buffer. An antibody amplifier™ was used for staining. A HRP-conjugated anti-mouse secondary was used at 1/10 dilution.

  • Overlay histogram showing HeLa cells stained with ab13537 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13537, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537) at 1/1000 dilution + 0.1ug mouse recombinant protein. Additional lower band is probably degraded or in other ways modified Dnmt1.

    Predicted band size : 183 kDa
  • All lanes : Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537) at 1 µg/ml

    Lane 1 : HCT116 whole cell lysate
    Lane 2 : HCT116 DNMT1 KO whole cell lysate
    Lane 3 : HCT116 DNMT3b KO whole cell lysate

    Secondary
    HRP conjugated Sheep anti-mouse IgG
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 183 kDa


    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537)

This product has been referenced in:
  • Chen M  et al. DNA methyltransferase inhibitor, zebularine, delays tumor growth and induces apoptosis in a genetically engineered mouse model of breast cancer. Mol Cancer Ther 11:370-82 (2012). WB . Read more (PubMed: 22203734) »
  • Leonard S  et al. Epigenetic and Transcriptional Changes Which Follow Epstein-Barr Virus Infection of Germinal Center B Cells and Their Relevance to the Pathogenesis of Hodgkin's Lymphoma. J Virol 85:9568-77 (2011). Read more (PubMed: 21752916) »

See all 15 Publications for this product

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Application Western blot
Sample Human Cell lysate - whole cell (lung carcinoma)
Loading amount 30 µg
Specification lung carcinoma
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Jan 18 2011

Application Western blot
Sample Human Cell lysate - whole cell (BT549)
Loading amount 40 µg
Specification BT549
Gel Running Conditions Reduced Denaturing (3-8% Tris-Acetate gel)
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Verified customer

Submitted Nov 18 2010

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (HeLa cytosolic and nuclear extracts)
Total protein in input 500 µg
Specification HeLa cytosolic and nuclear extracts
Immuno-precipitation step Protein G
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Verified customer

Submitted Nov 02 2009

Application Western blot
Sample Human Cell lysate - whole cell (HeLa cytosolic and nuclear extracts)
Loading amount 50 µg
Specification HeLa cytosolic and nuclear extracts
Gel Running Conditions Non-reduced Denaturing (8% gel)
Blocking step Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Verified customer

Submitted Jun 26 2009

Application Western blot
Sample Human Cell lysate - whole cell (Colon Carcinoma)
Loading amount 500000 cells
Specification Colon Carcinoma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5
Username

Verified customer

Submitted Mar 07 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"