Recombinant Anti-Dnmt3a antibody [EPR18455-94] (ab227823)
![Western blot - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)](https://www.abcam.com/ps/products/227/ab227823/Images/ab227823-389646-anti-dnmt3a-antibody-epr18455-94-western-blot-human-lysate.jpg "Western blot - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18455-94] to Dnmt3a
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Dnmt3a antibody [EPR18455-94]
See all Dnmt3a primary antibodies -
Description
Rabbit monoclonal [EPR18455-94] to Dnmt3a -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: ChIP,Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HEK-293T, HeLa and NIH/3T3 whole cell lysates; rat brain lysate. IHC-P: Human breast carcinoma and placenta tissue; mouse colon tissue; rat colon tissue.
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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
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
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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, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18455-94 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227823 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 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 125 kDa (predicted molecular weight: 102 kDa).
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| IHC-P |
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 125 kDa (predicted molecular weight: 102 kDa). |
|
IHC-P
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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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
modificationsSumoylated; 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
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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
see all -
Alternative names
- DNA (cytosine 5) methyltransferase 3 alpha antibody
- DNA (cytosine 5) methyltransferase 3A antibody
- DNA (cytosine-5)-methyltransferase 3A antibody
see all
Images
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Western blot - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)All lanes : Anti-Dnmt3a antibody [EPR18455-94] (ab227823) at 1/1000 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 kDaLanes 1-4: Merged signal (red and green). Green - ab227823 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.
ab227823 Anti-Dnmt3a antibody [EPR18455-94] 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. ab227823 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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.
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![Western blot - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)](https://www.abcam.com/ps/products/227/ab227823/Images/ab227823-5-227823WB1.jpg)
Western blot - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)All lanes : Anti-Dnmt3a antibody [EPR18455-94] (ab227823) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Dnmt3a knockout HAP1 whole cell lysate
Lane 3 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 102 kDaBlocking and dilution buffer: 5% NFDM/TBST.
ab227823 was shown to specifically react with wild-type HAP1 cells as signal was lost in Dnmt3a knockout cells. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab227823 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
The observed bands ranging from 75~125kDa represent different Dnmt3a isoforms.
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![Western blot - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)](https://www.abcam.com/ps/products/227/ab227823/Images/ab227823-6-227823WB2.jpg)
Western blot - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)Anti-Dnmt3a antibody [EPR18455-94] (ab227823) at 1/1000 dilution + Rat brain lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 102 kDa
Observed band size: 125 kDa why is the actual band size different from the predicted?
Exposure time: 26 secondsBlocking and dilution buffer: 5% NFDM/TBST.
The observed bands ranging from 75~125kDa represent different Dnmt3a isoforms.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)](https://www.abcam.com/ps/products/227/ab227823/Images/ab227823-4-227823IHC4.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the rat colon (PMID: 24837369) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.
The section was incubated with ab227823 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)](https://www.abcam.com/ps/products/227/ab227823/Images/ab227823-3-227823IHC3.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the mouse colon (PMID: 24837369) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.
The section was incubated with ab227823 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)](https://www.abcam.com/ps/products/227/ab227823/Images/ab227823-2-227823IHC2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human placenta (PMID: 22413869) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.
The section was incubated with ab227823 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)](https://www.abcam.com/ps/products/227/ab227823/Images/ab227823-1-227823IHC1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] (ab227823)Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human breast carcinoma (PMID: 24464625) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.
The section was incubated with ab227823 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
-
![Anti-Dnmt3a antibody [EPR18455-94] (ab227823)](https://www.abcam.com/ps/products/227/ab227823/Images/ab227823-8-benefits-of-recombinant-antibodies.png)
Anti-Dnmt3a antibody [EPR18455-94] (ab227823)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab227823 has been referenced in 1 publication.
- Zhou L et al. Melatonin inhibits lipid accumulation to repress prostate cancer progression by mediating the epigenetic modification of CES1. Clin Transl Med 11:e449 (2021). PubMed: 34185414