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    products/primary-antibodies/dnmt3a-antibody-epr18455-ab188470.pdf

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Epigenetics and Nuclear Signaling DNA methylation DNMTs
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Dnmt3a antibody [EPR18455] (ab188470)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (3) Submit a question References (20)

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Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Flow Cytometry (Intracellular) - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3a antibody [EPR18455] (ab188470)
  • Anti-Dnmt3a antibody [EPR18455] (ab188470)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18455] to Dnmt3a
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

APC Carrier Free HRP PE

You may also be interested in

Primary
Product image
Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (ab232391)
Assay
Product image
Cardiolipin Assay Kit (ab241036)
Knockout
Product image
Human DNMT3A knockout HeLa cell line (ab261793)

View more associated products

Overview

  • Product name

    Anti-Dnmt3a antibody [EPR18455]
    See all Dnmt3a primary antibodies
  • Description

    Rabbit monoclonal [EPR18455] to Dnmt3a
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HEK-293 and C6 cell lysates; Rat brain and heart lysates. ICC/IF: HeLa cells. IHC-P: Human placenta, mouse testis, rat spleen tissue
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18455
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA methylation
    • DNMTs

Associated products

  • Alternative Versions

    • Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (ab232391)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human DNMT3A knockout HeLa cell line (ab261793)
  • KO cell lysates

    • Human DNMT3A knockout HeLa cell lysate (ab257128)
  • Recombinant Protein

    • Recombinant human Dnmt3a protein (ab170408)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab188470 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB (2)
1/2000. Detects a band of approximately 130 kDa (predicted molecular weight: 102 kDa).
ICC/IF
1/1000.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/2000. Detects a band of approximately 130 kDa (predicted molecular weight: 102 kDa).
ICC/IF
1/1000.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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.
  • Target information above from: UniProt accession Q9Y6K1 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1788 Human
    • Entrez Gene: 13435 Mouse
    • Entrez Gene: 444984 Rat
    • Omim: 602769 Human
    • SwissProt: Q9Y6K1 Human
    • SwissProt: O88508 Mouse
    • SwissProt: Q1LZ53 Rat
    • Unigene: 515840 Human
    • Unigene: 5001 Mouse
    • Unigene: 92659 Rat
    see all
  • 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

  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    All lanes : Anti-Dnmt3a antibody [EPR18455] (ab188470) at 1/2000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : Dnmt3a knockout HeLa cell lysate
    Lane 3 : HEK-293 cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 102 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab188470 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab188470 Anti-Dnmt3a antibody [EPR18455] was shown to specifically react with Dnmt3a in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261793 (knockout cell lysate ab257128) was used. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab188470 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] (ab188470)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue sections labeling Dnmt3a with purified ab188470 at 1/2000 (0.409 µg/ml). Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain.

  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    All lanes : Anti-Dnmt3a antibody [EPR18455] (ab188470) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : Dnmt3a knockout HAP1 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Mouse brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 102 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab188470 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab188470 was shown to recognize Dnmt3a when Dnmt3a knockout samples were used, along with additional cross-reactive bands. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab188470 and ab8245 (loading control to GAPDH) were diluted to 1/5000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry (Intracellular) - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Flow Cytometry (Intracellular) - Anti-Dnmt3a antibody [EPR18455] (ab188470)

    Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Dnmt3a with purified ab188470 at 1/80 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

     

     

  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    All lanes : Anti-Dnmt3a antibody [EPR18455] (ab188470) at 1/5000 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : Mouse skin tissue lysate
    Lane 3 : Rat skin tissue lysate
    Lane 4 : Mouse brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 102 kDa
    Observed band size: 125 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab188470 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

     

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab188470 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] (ab188470)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue sections labeling Dnmt3 with purified ab188470 at 1/2000 (0.409 µg/ml). Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] (ab188470)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling Dnmt3a with purified ab188470 at 1/2000 (0.409 µg/ml). Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain.

  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Anti-Dnmt3a antibody [EPR18455] (ab188470) at 1/10000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 102 kDa
    Observed band size: 130 kDa why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed MW is consistent with what has been described in the literature (J Biol Chem. 2002. 277, 38746-38754. PMID: 2138111).

  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Anti-Dnmt3a antibody [EPR18455] (ab188470) at 1/10000 dilution + HEK-293 (Human epithelial cells from embryonic kidney) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 102 kDa
    Observed band size: 130 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed MW is consistent with what has been described in the literature (J Biol Chem. 2002. 277, 38746-38754. PMID: 2138111).

  • Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Western blot - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    All lanes : Anti-Dnmt3a antibody [EPR18455] (ab188470) at 1/2000 dilution

    Lane 1 : Rat brain lysate
    Lane 2 : Rat heart lysate
    Lane 3 : C6 (Rat glial tumor cells) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 102 kDa
    Observed band size: 130 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3a antibody [EPR18455] (ab188470)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Dnmt3a with ab188470 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab188470 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Anti-Dnmt3a antibody [EPR18455] (ab188470)
    Anti-Dnmt3a antibody [EPR18455] (ab188470)

Protocols

  • Flow cytometry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (20)

Publishing research using ab188470? Please let us know so that we can cite the reference in this datasheet.

ab188470 has been referenced in 20 publications.

  • Small L  et al. Ablation of DNA-methyltransferase 3A in skeletal muscle does not affect energy metabolism or exercise capacity. PLoS Genet 17:e1009325 (2021). PubMed: 33513138
  • Wang A  et al. MicroRNA-29a inhibits cell proliferation and arrests cell cycle by modulating p16 methylation in cervical cancer. Oncol Lett 21:272 (2021). PubMed: 33717269
  • Pi T  et al. High Methionine Diet-Induced Alzheimer's Disease like Symptoms Are Accompanied by 5-Methylcytosine Elevated Levels in the Brain. Behav Neurol 2021:6683318 (2021). PubMed: 33880134
  • Zhang JJ  et al. DNMT3a in the hippocampal CA1 is crucial in the acquisition of morphine self-administration in rats. Addict Biol 25:e12730 (2020). PubMed: 30950138
  • Chen Q  et al. miR-30a-3p inhibits the proliferation of liver cancer cells by targeting DNMT3a through the PI3K/AKT signaling pathway. Oncol Lett 19:606-614 (2020). PubMed: 31897176
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-Dnmt3a antibody [EPR18455]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS)
Gel Running Conditions
Reduced Denaturing (15)
Loading amount
50 µg
Specification
U2OS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 18 2022

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Dnmt3a antibody [EPR18455]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Astatotilapia burtoni Tissue sections (thalamus/hypothalamus)
Antigen retrieval step
None
Permeabilization
Yes - 0.2% TritonX
Specification
thalamus/hypothalamus
Blocking step
10% normal serum 1% BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Sebastian Alvarado

Verified customer

Submitted Dec 18 2015

Western blot abreview for Anti-Dnmt3a antibody [EPR18455]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Astatotilapia burtoni Cell lysate - whole cell (Brain and gonads)
Gel Running Conditions
Reduced Denaturing (5% acrylamide gel)
Loading amount
25 µg
Specification
Brain and gonads
Blocking step
Milk as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Sebastian Alvarado

Verified customer

Submitted Aug 07 2015

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