Overview

  • Product nameAnti-Dnmt3a antibody - ChIP Grade
    See all Dnmt3a primary antibodies
  • Description
    Rabbit polyclonal to Dnmt3a - ChIP Grade
  • SpecificityThis antibody detects DNA methyltransferase 3a (Dnmt3a) from human and mouse tissues and cells as well as recombinant human Dnmt3a. This antibody does not detect full length recombinant human Dnmt3b or Dnmt1.
  • Tested applicationsSuitable for: IHC-P, IP, WB, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Drosophila melanogaster
  • Immunogen

    Synthetic peptide corresponding to Human Dnmt3a aa 86-100.
    Sequence:

    DLLPNGDLEKRSEPQ


    (Peptide available as ab4921)

  • Positive control
    • HeLa cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab2850 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
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 101.5 kDa).Can be blocked with Dnmt3a peptide (ab4921).
ChIP Use at an assay dependent concentration. PubMed: 19620278

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. It modifies DNA in a non-processive manner and also methylates non-CpG sites. May preferentially methylate DNA linker between 2 nucleosomal cores and is inhibited by histone H1. Plays a role in paternal and maternal imprinting. Required for methylation of most imprinted loci in germ cells. Acts as a transcriptional corepressor for ZNF238. Can actively repress transcription through the recruitment of HDAC activity.
  • Tissue specificityHighly expressed in fetal tissues, skeletal muscle, heart, peripheral blood mononuclear cells, kidney, and at lower levels in placenta, brain, liver, colon, spleen, small intestine and lung.
  • 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; sumoylation disrupts the ability to interact with histone deacetylases (HDAC1 and HDAC2) and repress transcription.
  • Cellular localizationNucleus. Cytoplasm. Accumulates in the major satellite repeats at pericentric heterochromatin.
  • Information by UniProt
  • Database links
  • Alternative names
    • DNA (cytosine 5) methyltransferase 3 alpha antibody
    • DNA (cytosine 5) methyltransferase 3A antibody
    • DNA (cytosine-5)-methyltransferase 3A antibody
    • DNA cytosine methyltransferase 3A2 antibody
    • DNA methyltransferase 3 alpha antibody
    • DNA methyltransferase 3a antibody
    • DNA methyltransferase HsaIIIA antibody
    • DNA MTase HsaIIIA antibody
    • DNM3A_HUMAN antibody
    • DNMT 3a antibody
    • DNMT antibody
    • Dnmt3a antibody
    • DNMT3A2 antibody
    • M.HsaIIIA antibody
    • MCMT antibody
    • OTTHUMP00000201149 antibody
    • TBRS antibody
    see all

Anti-Dnmt3a antibody - ChIP Grade images



  • Predicted band size : 101.5 kDa

    Image from Nucleic Acids Res. 2014 Jan; 42(2): 804–821. Fig 5D, doi: 10.1093/nar/gkt920

    Western blot analysis of DNMT3a in HeLa silenced cells. Total cell extracts were prepared 48 h after electroporation and analyzed by immunoblot with ab2850.

  • DNMT3a is recruited to the DSB early during repair and is necessary for generation of L cells. HeLa cells were transfected with I-SceI (a meganuclease) and 24 h, 48 h or 7 days later, were fixed, collected, chromatin-extracted and subjected to ChIP analysis with ab2820. Figure 5B shows that DNMT3a is recruited to the I-SceI site 24 h after the onset of DSB formation and rapidly disappeared (48 h).

    Cells were fixed by adding formaldehyde directly in the culture medium to a final concentration of 1% for 10 min at RT and washed twice using ice cold PBS containing 1× protease inhibitor cocktail and 1 mM PMSF. Fixed cells were harvested and the pellet was resuspended in 200 µl of SDS Lysis Buffer. After 10 min incubation on ice, the lysates were sonicated. Sonicated samples were centrifuged and supernatants diluted 10-fold in the ChIP Dilution Buffer. An aliquot (1/50) of sheared chromatin was further treated with proteinase K, phenol/chloroform extracted and precipitated to determine DNA concentration and shearing efficiency (input DNA).

    Sheared chromatin was precleared,divided in two aliquots and incubated at 4°C for 16 h with 20 µl of protein-A/G agarose and 2 µg of ab2850 and nonimmune IgG. Agarose beads were washed and immunoprecipitated DNA was recovered and subjected to qPCR.



  • Predicted band size : 101.5 kDa
    Western blot of Dnmt3a on P19 cell nuclear extract using ab2850.

References for Anti-Dnmt3a antibody - ChIP Grade (ab2850)

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) »
  • Bahar Halpern K  et al. Paradoxical Role of DNA Methylation in Activation of FoxA2 Gene Expression during Endoderm Development. J Biol Chem 289:23882-92 (2014). ChIP . Read more (PubMed: 25016019) »

See all 11 Publications for this product

Product Wall

Application Western blot
Sample Rat Cell lysate - whole cell (Gonad)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 75 µg
Specification Gonad
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Nov 11 2015

Application ChIP
Sample Human Cell lysate - other (Epithelial cells)
Negative control IgG
Specification Epithelial cells
Detection step Real-time PCR
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Positive control Input DNA
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Submitted Sep 30 2015

Application ChIP
Detection step Real-time PCR
Sample Rat Tissue lysate - whole (Gonad)
Specification Gonad
Negative control IgG
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Positive control Input DNA
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Submitted Jul 07 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako antigen retrieval solution
Sample Marmoset (common) Tissue sections (Adult testis)
Specification Adult testis
Permeabilization No
Fixative Formaldehyde
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Submitted Jan 21 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

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