Anti-Dnmt3a抗体[64B1446] - ChIP Grade (ab13888)

概述

  • 产品名称Anti-Dnmt3a抗体[64B1446] - ChIP Grade
    参阅全部 Dnmt3a 一抗
  • 描述
    小鼠单克隆抗体[64B1446] to Dnmt3a - ChIP Grade
  • 经测试应用适用于: WB, IHC-P, ChIP, Flow Cyt, ICC/IF, ICCmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Bacteria expressed HIS-tag recombinant mouse Dnmt3a.

  • 表位Found to lie near the C terminus (a.a. 705-908).
  • 阳性对照

性能

应用

Our Abpromise guarantee covers the use of ab13888 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 2 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 102 kDa). Use at a concentration of 2 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 102 kDa). Dnmt3a Transfected and Untransfected Cell Lysate (ab90154) is the recommended positive control for WB.
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration.
ICC Use a concentration of 5 µg/ml.

靶标

  • 功能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.
  • 组织特异性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.
  • 序列相似性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.
  • 结构域The PWWP domain is essential for targeting to pericentric heterochromatin.
  • 翻译后修饰Sumoylated; sumoylation disrupts the ability to interact with histone deacetylases (HDAC1 and HDAC2) and repress transcription.
  • 细胞定位Nucleus. Cytoplasm. Accumulates in the major satellite repeats at pericentric heterochromatin.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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 [64B1446] - ChIP Grade 图像



  • Predicted band size : 102 kDa


    Western blot analysis for Dnmt3a using ab13888 at 2 ug/ml against 10 µg of 293 cell lysate transfected with either mouse Dnmt3a (lane A) or mouse Dnmt3b (lane B).
  • Immuno-staining of Dnmt3a using ab13888 at 5 ug/ml dilution on cultured HeLa cells. The nucleic staining is shown in most of the cells.

  • ab13888 (2µg/ml) staining Dnmt3a in human testis using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining in the seminal vesicles.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • ICC/IF image of ab13888 stainedHepG2 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 (ab13888, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) 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.

  • Overlay histogram showing HEK293 cells stained with ab13888 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab138889, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Anti-Dnmt3a antibody [64B1446] - ChIP Grade (ab13888)参考文献

This product has been referenced in:
  • Lachance G  et al. DNMT3a epigenetic program regulates the HIF-2a oxygen-sensing pathway and the cellular response to hypoxia. Proc Natl Acad Sci U S A 111:7783-8 (2014). Read more (PubMed: 24817692) »
  • Castro-Diaz N  et al. Evolutionally dynamic L1 regulation in embryonic stem cells. Genes Dev 28:1397-409 (2014). Human . Read more (PubMed: 24939876) »

See all 17 Publications for this product

Product Wall



The Dnmt3a antibody (ab13888) is a mouse IgG1 monoclonal antibody. We have two secondary FITC conjugate antibody on our catalog:

Goat Anti-Mouse IgG1 heavy chain (FITC) http://www.abcam.com/index.html?datasheet=97239 (or use the foll...

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If there is no other protocol then our standard protocols suggested on the product data sheet, the general protocols should be sufficient. However, I contacted the laboratory to get the protocol that was used doing the testing of the antibody. The prot...

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I can recommend the following secondary antibodies, which are enzyme conjugated to HRP that would be suitable for the detection of the three primary antibodies (Anti-Dnmt3b antibody [52A1018] - ChIP Grade (ab13604); Anti-Dnmt1 antibody [60B1220.1] - Ch...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (liver cancer)
Loading amount 10 µg
Specification liver cancer
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Oct 17 2012

Application Western blot
Sample Rat Cell lysate - whole cell (Cortical cells 15 DIV)
Loading amount 20 µg
Specification Cortical cells 15 DIV
Gel Running Conditions Reduced Denaturing (5)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jul 06 2012

Thank you for contacting Abcam.

It could be that ab13888, is recognizing a non-specific band in the cell lines that you are using, but you should still be seeing a band at about 100kDa.

To be able to help resolve this issue, could y...

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Thank you for contacting Abcam.

I have attached the ppt slide with a red arrow next to the bands that I think are the correct bands(they are the same that you have marked).

There are a few protocol changes that I would recommend to ...

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