Overview

  • Product name
    Anti-Dnmt3a antibody - ChIP Grade
    See all Dnmt3a primary antibodies
  • Description
    Rabbit polyclonal to Dnmt3a - ChIP Grade
  • Specificity
    This 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 applications
    Suitable 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

Associated products

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

  • Function
    Required 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 specificity
    Highly 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 similarities
    Belongs 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.
  • Domain
    The 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 localization
    Nucleus. 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:
  • Chen S  et al. Histone H2B monoubiquitination is a critical epigenetic switch for the regulation of autophagy. Nucleic Acids Res 45:1144-1158 (2017). WB ; Human . Read more (PubMed: 28180298) »
  • O'Leary VB  et al. Long non-coding RNA PARTICLE bridges histone and DNA methylation. Sci Rep 7:1790 (2017). WB ; Human . Read more (PubMed: 28496150) »

See all 27 Publications for this product

Product Wall

Application
Western blot
Sample
Rat Cell lysate - whole cell (HEK and NRK)
Gel Running Conditions
Reduced Denaturing (6)
Loading amount
20 µg
Specification
HEK and NRK
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Apr 28 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (dorsal root ganglion and spinal cord (P4))
Permeabilization
Yes - 0.2% Triton
Specification
dorsal root ganglion and spinal cord (P4)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Dr. Franziska Denk

Verified customer

Submitted Mar 22 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - whole cell (Breast cancer line)
Specification
Breast cancer line
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 14 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Breast cancer line)
Gel Running Conditions
Reduced Denaturing (12.5%)
Loading amount
100000 cells
Specification
Breast cancer line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Mar 07 2017

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

煜欽 林

Verified customer

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
Username

Abcam user community

Verified customer

Submitted Sep 23 2013

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

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