Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3522] to Dnmt1
- Suitable for: WB, IP, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Human
Product nameAnti-Dnmt1 antibody [EPR3522]
See all Dnmt1 primary antibodies
DescriptionRabbit monoclonal [EPR3522] to Dnmt1
Tested applicationsSuitable for: WB, IP, Flow Cyt, ICC/IFmore details
Unsuitable for: IHC-P
Species reactivityReacts with: Human
Synthetic peptide within Human Dnmt1 aa 1600-1700 (C terminal). The exact sequence is proprietary.
- HuT-78, Jurkat or 293T lysate HeLa cells
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
Our Abpromise guarantee covers the use of ab92314 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Predicted molecular weight: 183 kDa.|
|Flow Cyt||1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/50 - 1/100.|
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.
modificationsSumoylated; sumoylation increases activity.
- Information by UniProt
- ADCADN antibody
- AIM antibody
- CXXC finger protein 9 antibody
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: DNMT1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HEK293 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab92314 observed at 170 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab92314 was shown to specifically react with DNMT1 when DNMT1 knockout samples were used. Wild-type and DNMT1 knockout samples were subjected to SDS-PAGE. Ab92314 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 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.
Immunofluorescence staining of Jurkat cells with purified ab92314 at a working dilution of 1/2000, counter-stained with DAPI. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-rabbit (ab150077), used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.
Overlay histogram showing HeLa cells stained with ab92314 (red line). The cells were fixed with 80% 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 (ab92314, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
All lanes : Anti-Dnmt1 antibody [EPR3522] (ab92314) at 1/1000 dilution
Lane 1 : HuT-78 cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : 293T cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 183 kDa
ab92314 has been referenced in 5 publications.
- Molokie R et al. Oral tetrahydrouridine and decitabine for non-cytotoxic epigenetic gene regulation in sickle cell disease: A randomized phase 1 study. PLoS Med 14:e1002382 (2017). PubMed: 28880867
- Heuslein JL et al. DNA Methyltransferase 1-Dependent DNA Hypermethylation Constrains Arteriogenesis by Augmenting Shear Stress Set Point. J Am Heart Assoc 6:N/A (2017). PubMed: 29191807
- Goel S et al. CDK4/6 inhibition triggers anti-tumour immunity. Nature 548:471-475 (2017). WB ; Mouse . PubMed: 28813415
- Trejbalová K et al. Development of 5' LTR DNA methylation of latent HIV-1 provirus in cell line models and in long-term-infected individuals. Clin Epigenetics 8:19 (2016). WB . PubMed: 26900410
- Clements EG et al. DNMT1 modulates gene expression without its catalytic activity partially through its interactions with histone-modifying enzymes. Nucleic Acids Res 40:4334-46 (2012). WB . PubMed: 22278882