Overview

  • Product nameAnti-Dnmt3b antibody - ChIP Grade
    See all Dnmt3b primary antibodies
  • Description
    Rabbit polyclonal to Dnmt3b - ChIP Grade
  • SpecificityThis antibody detects DNA methyltransferase 3b (Dnmt3b) from human and mouse tissues and cells as well as recombinant human Dnmt3b. This antibody detects, to a lesser extent, full-length human recombinant Dnmt3a.
  • Tested applicationsSuitable for: ICC/IF, IP, IHC-P, WB, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse Dnmt3b aa 1-14.
    Sequence:

    MKGDSRHLNEEEGA


    (Peptide available as ab4922)

  • Positive control
    • P19 nuclear extracts. ChIP: PMID: 16357870 (Vire E et al Nature 2006 439:871) has used U2OS cells and primers specific for the hsMYT1 promoter. The primers are described in PMID: 15231737 (Kirmizis A et al Genes and Dev 2004 18:1592).

Properties

Applications

Our Abpromise guarantee covers the use of ab2851 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 Use a concentration of 1 µg/ml.
IP Use at an assay dependent concentration. PubMed: 16631596
IHC-P 1/100 - 1/1000.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 97.5 kDa).Can be blocked with Dnmt3b peptide (ab4922).
ChIP Use at an assay dependent concentration. PubMed: 17972916

Target

  • FunctionRequired 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. May preferentially methylates nucleosomal DNA within the nucleosome core region. May function as transcriptional co-repressor by associating with CBX4 and independently of DNA methylation. Seems to be involved in gene silencing (By similarity). In association with DNMT1 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. Isoforms 4 and 5 are probably not functional due to the deletion of two conserved methyltransferase motifs.
  • Tissue specificityUbiquitous; highly expressed in fetal liver, heart, kidney, placenta, and at lower levels in spleen, colon, brain, liver, small intestine, lung, peripheral blood mononuclear cells, and skeletal muscle. Isoform 1 is expressed in all tissues except brain, skeletal muscle and PBMC, 3 is ubiquitous, 4 is expressed in all tissues except brain, skeletal muscle, lung and prostate and 5 is detectable only in testis and at very low level in brain and prostate.
  • Involvement in diseaseDefects in DNMT3B are a cause of immunodeficiency-centromeric instability-facial anomalies syndrome (ICF) [MIM:242860]. ICF is a rare autosomal recessive disorder characterized by a variable immunodeficiency, mild facial anomalies, and centromeric heterochromatin instability involving chromosomes 1, 9, and 16. ICF is biochemically characterized by hypomethylation of CpG sites in some regions of heterochromatin.
  • Sequence similaritiesBelongs 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.
  • DomainThe PWWP domain is essential for targeting to pericentric heterochromatin.
  • Post-translational
    modifications
    Sumoylated.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Cytosine 5methyltransferase 3B antibody
    • DNA antibody
    • DNA (cytosine 5) methyltransferase 3 beta antibody
    • DNA (cytosine 5)-methyltransferase 3B antibody
    • DNA (cytosine-5)-methyltransferase 3B antibody
    • DNA methyltransferase HsaIIIB antibody
    • DNA MTase HsaIIIB antibody
    • DNM3B_HUMAN antibody
    • Dnmt3b antibody
    • EC 2.1.1.37 antibody
    • ICF antibody
    • ICF1 antibody
    • M.HsaIIIB antibody
    • MGC124407 antibody
    • RP23-89H14.3 antibody
    see all

Anti-Dnmt3b antibody - ChIP Grade images

  • ab2851 labellling Dnmt3b in the nucleus of Mouse brain tissue (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS. Tissue sections were incubated with primary antibody (1:200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • ab2851 labellling Dnmt3b in the nucleus of Human testis tissue (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS. Tissue sections were incubated with primary antibody (1:500 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • ab2851 labellling Dnmt3b in the nucleus and cytoplasm of Human breast carcinoma (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS. Tissue sections were incubated with primary antibody (1:500 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • All lanes : Anti-Dnmt3b antibody - ChIP Grade (ab2851) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : A549 cell lysate
    Lane 3 : NIH-3T3 cell lysate

    Lysates/proteins at 25 µg per lane.


    Predicted band size : 97.5 kDa
    Observed band size : 97 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab2851 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2851, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-Dnmt3b antibody - ChIP Grade (ab2851) at 1 µg/ml

    Lane 1 : A498 (Human Kidney Carcinoma) Whole Cell Lysate
    Lane 2 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 97.5 kDa
    Observed band size : 97.5 kDa
    Additional bands at : 32 kDa,46 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes

References for Anti-Dnmt3b antibody - ChIP Grade (ab2851)

This product has been referenced in:
  • Shen W  et al. Epigenetic modification of the leptin promoter in diet-induced obese mice and the effects of N-3 polyunsaturated Fatty acids. Sci Rep 4:5282 (2014). ChIP ; Mouse . Read more (PubMed: 24923522) »
  • Castro-Diaz N  et al. Evolutionally dynamic L1 regulation in embryonic stem cells. Genes Dev 28:1397-409 (2014). Human . Read more (PubMed: 24939876) »

See all 22 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Muscle)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA 20 min at 97ºC in a PT link
Permeabilization No
Specification Muscle
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Sep 24 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Cardiomyocytes)
Specification Cardiomyocytes
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
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Submitted Sep 18 2015

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Brain)
Permeabilization Yes - Triton x-100, 0.01%
Specification Brain
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
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Submitted Jun 29 2015

Application Western blot
Sample Mouse Cell lysate - whole cell (Cardiomyocytes)
Gel Running Conditions Reduced Denaturing (10% PAGE)
Loading amount 25 µg
Specification Cardiomyocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Jun 29 2015

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Thank you for contacting us and taking the time to send the questionnaire. We really appreciate all the details provided as well as the images sent, that help us to better understand the problem. All of our products are covered by our Abpromise guaran...

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Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Hepatoma cells)
Specification Hepatoma cells
Fixative Formaldehyde
Permeabilization Yes - 0.2% triton X-100
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Jul 23 2008

A brief literature search revealed that PMID: 16357870 (Vire E et al Nature 2006 439:871) has used U2OS cells and primers specific for the hsMYT1 promoter. The primers are described in PMID: 15231737 (Kirmizis A et al Genes and Dev 2004 18:1592). I ...

Read More

Thank you for getting back in touch with me. To clarify. A predicted molecular weight of 97KDa for Dnmt3b is derived from SwissProt reference Q9UBC3. This is purely based on the protein sequence and a prediction. ab2851 detects a band of approximate...

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I have been advised by the source of ab2851 that it would be most important to run a positive control. In the lab recombinant DNMT3b has been used, however it is reported that DNMT3b is highly expressed in P19 (murine EC) cells when compared with human...

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1-10 of 17 Abreviews or Q&A

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