Overview

  • Product name

    Anti-Dnmt3a (phospho S390 + S393) antibody
    See all Dnmt3a primary antibodies
  • Description

    Rabbit polyclonal to Dnmt3a (phospho S390 + S393)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Rat, Sheep, Rabbit, Horse, Cow, Dog, Pig, Chimpanzee, Macaque monkey, Gorilla, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human Dnmt3a aa 350-450 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab130812, ab130813, ab133980)

  • Positive control

    • This antibody gave a positive signal in HEK293 whole cell lysate. This antibody gave a positive signal in the following methanol fixed cell lines: HepG2.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
    Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab87763 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 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 102 kDa).

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

Images

  • All lanes : Anti-Dnmt3a (phospho S390 + S393) antibody (ab87763) at 1 µg/ml

    Lane 1 : HEK293 (Human) Whole Cell Lysate (ab52256)
    Lane 2 : HEK293 (Human) Whole Cell Lysate (ab52256) with Phospho S390 + S393 peptide at 1 µg/ml
    Lane 3 : HEK293 (Human) Whole Cell Lysate (ab52256) with Phospho S390 peptide at 1 µg/ml
    Lane 4 : HEK293 (Human) Whole Cell Lysate (ab52256) with Phospho S393 peptide at 1 µg/ml
    Lane 5 : HEK293 (Human) Whole Cell Lysate (ab52256) with Non modified control peptide at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : 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: 102 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 39 kDa, 52 kDa, 68 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes
  • Lysates from U2OS WT cells were loaded as input. The western blot was probed with anti Dnmt3a (phospho S390 + S393) antibody (ab87763) either in the presence or absence of double modified peptide phosphorylated on S390 and S393 as indicated in the figure. Detected phospho Dnmt3a is indicated by the arrow. Please note: the observed band migrates at a higher MW than predicted probably because of phosphorylation. This data was provided by an anonymous collaborator.
  • ICC/IF image of ab87763 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 ab87763 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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

This product has been referenced in:

  • Deplus R  et al. Regulation of DNA methylation patterns by CK2-mediated phosphorylation of Dnmt3a. Cell Rep 8:743-53 (2014). WB ; Mouse, Human . Read more (PubMed: 25066127) »
See 1 Publication for this product

Customer reviews and Q&As

Answer

I am sorry to confirm that as far as we are aware, ab87763 has never been tested in samples from mouse. All tested species and applications covered by the guarantee are listed on the individual datasheets, and these are updated as soon as any new information is brought to our attention.

I have checked the alignment of the immunogen for the antibody with the mouse protein sequence, and this is 71%. We recommend that there should be 85% alignment to predict that an antibody will detect in another species. However, at 71%, there could still be some crossreactivity.

I also checked for the conservation of the phosphosites between human and mouse (http://www.phosphosite.org/proteinAction.do?id=20751&showAllSites=true), they are slightly shifted but this is also reported for the rat phosphosites, a species predicted to react with the above mentioned antibody.

If you would like to test ab87763 in mouse, please do not hesitate to contact me again by replying to this message prior to the purchase as you may be eligible for our AbTrial testing discount program. More information regarding this offer can be found at: www.abcam.com/abtrial.

Alternatively, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews


We are also now offering custom RabMAb antibody services through Epitomics – an Abcam company. If you are continuing to have difficulties finding an antibody to detect these phospho sites on the mouse protein, you may be interested in learning more. We can connect you with one of our project managers to discuss your antibody needs.

www.abcam.com/custom-support
email: service@abcam.com
Phone: (877) 772-2622 Press option 3

Read More

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