Overview

  • Product nameAnti-Dnmt3a antibody
    See all Dnmt3a primary antibodies
  • Description
    Rabbit polyclonal to Dnmt3a
  • Specificityab23565 detects His-tagged-recombinant dnmt3a at 120 kDa. However, ab23565 does not appear to recognise dnmt3a in HeLa cell lines.
  • Tested applicationsICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Mouse Dnmt3a.

    (Peptide available as ab26339.)

  • Positive control
    • This antibody gave a positive signal in the following Whole Cell Lysates: Jurkat, Hek293

Properties

  • FormLiquid
  • Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.1% Sodium Azide
    Constituents: PBS, pH 7.6
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Clonality Polyclonal
  • IsotypeIgG
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab23565 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 10 µg/ml.
IHC-P 1/300. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Can be blocked with Human Dnmt3a peptide (ab26339).

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 3 alpha antibody
    • DNA (cytosine 5) methyltransferase 3A antibody
    • DNA (cytosine-5)-methyltransferase 3A antibody
    • DNA cytosine methyltransferase 3A2 antibody
    • DNA methyltransferase 3a antibody
    • DNA methyltransferase HsaIIIA antibody
    • DNA MTase HsaI antibody
    • DNA MTase HsaIIIA antibody
    • DNM3A_HUMAN antibody
    • DNMT 3a antibody
    • DNMT antibody
    • Dnmt3a antibody
    • DNMT3A2 antibody
    • M HsaIIIA antibody
    • M.HsaIIIA antibody
    • MCMT antibody
    • Methyl CpG binding domain protein 3a antibody
    • OTTHUMP00000201149 antibody
    see all

Anti-Dnmt3a antibody images

  • ab23565 staining Dnmt3a in mouse lung tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 25°C; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/300) for 24 hours at 4°C. A HRP-conjugated anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • Anti-Dnmt3a antibody (ab23565) at 1 µg/ml + dnmt3a recombinant protein

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090)

    Predicted band size : 120 kDa
    Observed band size : 120 kDa
  • All lanes : Anti-Dnmt3a antibody (ab23565) at 1/500 dilution

    Lane 1 : 40ug mouse brain tissue (transgenic mouse negative for Dnmt3a) - whole lysate
    Lane 2 : 40ug mouse spleen tissue (higher than normal levels of Dnmt3a) - whole lysate

    Secondary
    Goat anti-rabbit HRP
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 120 kDa
    Observed band size : 84 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 78 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview

    See Abreview

  • All lanes : Anti-Dnmt3a antibody (ab23565) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 120 kDa
    Observed band size : 105 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 65 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 90 seconds
    The band seen at 105 kDa is consistent with the banding pattern observed  for other commercially available antibodies to Dnmt3a.
  • ICC/IF image of ab23565 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab23565, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

References for Anti-Dnmt3a antibody (ab23565)

This product has been referenced in:
  • Dunn J  et al. Flow-dependent epigenetic DNA methylation regulates endothelial gene expression and atherosclerosis. J Clin Invest 124:3187-99 (2014). WB ; Mouse . Read more (PubMed: 24865430) »
  • Jefferson WN  et al. Persistently altered epigenetic marks in the mouse uterus after neonatal estrogen exposure. Mol Endocrinol 27:1666-77 (2013). WB ; Mouse . Read more (PubMed: 24002655) »

See all 6 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Based Ph 6.0
Sample Baboon Tissue sections (Endometrium)
Specification Endometrium
Permeabilization No
Fixative 10% Buffered Formalin
Username

Mark Olson

Verified customer

Submitted Oct 11 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step Heat mediated
Sample Mouse Tissue sections (Lung)
Specification Lung
Permeabilization No
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 18 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

Zachary Yu-Ching Lin

Verified customer

Submitted Jan 21 2014

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Antigen retrieval step None
Sample Rat Tissue sections (Brain)
Specification Brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Miss. Sarah Rudzinskas

Verified customer

Submitted Jan 17 2014

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (8-16)
Sample Rat Tissue lysate - whole (Medial Amygdala)
Specification Medial Amygdala
Blocking step BSA as blocking agent for 36 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Username

Miss. Sarah Rudzinskas

Verified customer

Submitted Jan 15 2014

Thank you for contacting us.

The immunogen peptide is a synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Mouse Dnmt3a.


I hope this information is helpful to you. Please do not hesitate to contact u...

Read More
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Dorsal root ganglion)
Specification Dorsal root ganglion
Fixative Paraformaldehyde
Permeabilization Yes - 3% Triton-X in PBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Username

Dr. Y Tao

Verified customer

Submitted Nov 07 2011

I am very pleased to hear you would like to accept our offer and test ab23565 in rat. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for rat and include this code in the “...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Spleen)
Loading amount 40 µg
Specification Spleen
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Username

Abcam user community

Verified customer

Submitted Nov 06 2006

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