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

Overview

  • Product nameAnti-Dnmt1 [60B1220.1] antibody - 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 control
    • Human 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 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 Use at an assay dependent dilution.
ChIP Use at an assay dependent dilution.
Flow Cyt 1/100.
WB Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 183 kDa).
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
    • 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 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

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

This product has been referenced in:
  • Kiianitsa K & Maizels N A rapid and sensitive assay for DNA-protein covalent complexes in living cells. Nucleic Acids Res 41:e104 (2013). WB ; Human . Read more (PubMed: 23519618) »
  • Wong YF  et al. Prolonged treatment with DNMT inhibitors induces distinct effects in promoters and gene-bodies. PLoS One 8:e71099 (2013). WB ; Human . Read more (PubMed: 23940695) »

See all 21 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (6%)
Sample Mouse Cell lysate - whole cell (Mouse N2A neuroblastoma)
Specification Mouse N2A neuroblastoma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Apr 03 2014

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (6%)
Sample Human Cell lysate - whole cell (hek 293)
Specification hek 293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Apr 02 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step Enzymatic
Sample Rat Tissue sections (GONAD)
Specification GONAD
Permeabilization No
Fixative Formaldehyde
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Submitted Sep 23 2013

Application Immunoprecipitation
Immuno-precipitation step Protein A
Sample Human Cell lysate - whole cell (fibroblasts)
Specification fibroblasts
Total protein in input 100000 cells
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Submitted Aug 19 2013

Application Western blot
Loading amount 1e+006 cells
Gel Running Conditions Reduced Denaturing (12 %)
Sample Human Cell lysate - other (fibroblasts)
Specification fibroblasts
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Aug 14 2013

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