Anti-Dnmt3b抗体[52A1018] - ChIP Grade (ab13604)

概述

  • 产品名称Anti-Dnmt3b抗体[52A1018] - ChIP Grade
    参阅全部 Dnmt3b 一抗
  • 描述
    小鼠单克隆抗体[52A1018] to Dnmt3b - ChIP Grade
  • 特异性Does not cross-react with Dnmt3a.
  • 经测试应用适用于: ChIP, IHC-P, WB, IP, ICC/IF, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    His-tagged recombinant protein (Mouse).

  • 阳性对照
    • P19 nuclear extract

性能

应用

Our Abpromise guarantee covers the use of ab13604 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
ChIP Use at an assay dependent concentration. PubMed: 15548683
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 110 kDa.
IP Use a concentration of 1 - 2 µg/ml.
ICC/IF Use a concentration of 1 - 2 µg/ml.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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. May preferentially methylates nucleosomal DNA within the nucleosome core region. May function as transcriptional co-repressor by associating with CBX4 and independently of DNA methylation. Seems to be involved in gene silencing (By similarity). In association with DNMT1 and via the recruitment of CTCFL/BORIS, involved in activation of BAG1 gene expression by modulating dimethylation of promoter histone H3 at H3K4 and H3K9. Isoforms 4 and 5 are probably not functional due to the deletion of two conserved methyltransferase motifs.
  • 组织特异性Ubiquitous; highly expressed in fetal liver, heart, kidney, placenta, and at lower levels in spleen, colon, brain, liver, small intestine, lung, peripheral blood mononuclear cells, and skeletal muscle. Isoform 1 is expressed in all tissues except brain, skeletal muscle and PBMC, 3 is ubiquitous, 4 is expressed in all tissues except brain, skeletal muscle, lung and prostate and 5 is detectable only in testis and at very low level in brain and prostate.
  • 疾病相关Defects in DNMT3B are a cause of immunodeficiency-centromeric instability-facial anomalies syndrome (ICF) [MIM:242860]. ICF is a rare autosomal recessive disorder characterized by a variable immunodeficiency, mild facial anomalies, and centromeric heterochromatin instability involving chromosomes 1, 9, and 16. ICF is biochemically characterized by hypomethylation of CpG sites in some regions of heterochromatin.
  • 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.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Cytosine 5methyltransferase 3B antibody
    • DNA antibody
    • DNA (cytosine 5) methyltransferase 3 beta antibody
    • DNA (cytosine 5)-methyltransferase 3B antibody
    • DNA (cytosine-5)-methyltransferase 3B antibody
    • DNA methyltransferase HsaIIIB antibody
    • DNA MTase HsaIIIB antibody
    • DNM3B_HUMAN antibody
    • Dnmt3b antibody
    • EC 2.1.1.37 antibody
    • ICF antibody
    • ICF1 antibody
    • M.HsaIIIB antibody
    • MGC124407 antibody
    • RP23-89H14.3 antibody
    see all

Anti-Dnmt3b antibody [52A1018] - ChIP Grade 图像

  • All lanes : Anti-Dnmt3b antibody [52A1018] - ChIP Grade (ab13604) at 2 µg/ml

    Lane 1 : Cell lysates prepared from mouse Dnmt3a transfected 293 cells.
    Lane 2 : Cell lysates prepared from mouse Dnmt3b transfected 293 cells

    Lysates/proteins at 10 µg per lane.


    Observed band size : 110 kDa (why is the actual band size different from the predicted?)
  • Ab13604 staining Human normal testis. Staining is localised to the nucleus.
    Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the DAKO 3-in-1 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • ICC/IF image of ab13604 stained HepG2 cells. The cells were 4% formaldehyde 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 (ab13604, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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 HepG2 cells stained with ab13604 (red line). The cells were fixed with 80% methanol (5 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 (ab13604, 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.

Anti-Dnmt3b antibody [52A1018] - ChIP Grade (ab13604)参考文献

This product has been referenced in:
  • Sheaffer KL  et al. DNA methylation is required for the control of stem cell differentiation in the small intestine. Genes Dev 28:652-64 (2014). Read more (PubMed: 24637118) »
  • Murgatroyd C & Spengler D Polycomb binding precedes early-life stress responsive DNA methylation at the Avp enhancer. PLoS One 9:e90277 (2014). ChIP ; Mouse . Read more (PubMed: 24599304) »

See all 29 Publications for this product

Product Wall

Application ChIP
Sample Human Cell lysate - other (Epithelial cells)
Negative control IgG
Specification Epithelial cells
Detection step Real-time PCR
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Positive control Input Control
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提交于 Sep 30 2015

Application ChIP
Detection step Real-time PCR
Sample Rat Tissue lysate - whole (Gonad)
Specification Gonad
Negative control IgG
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Positive control Input DNA
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提交于 Jul 07 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (8%)
Sample Human Cell lysate - whole cell (Jurkat cell line)
Specification Jurkat cell line
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
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提交于 Feb 14 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step Enzymatic
Sample Rat Tissue sections (GONAD)
Specification GONAD
Permeabilization No
Fixative Formaldehyde
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提交于 Sep 25 2013

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 20 µg
Specification liver cancer
Gel Running Conditions Reduced Denaturing (10%gel)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Dec 11 2012

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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)...

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Thank you for contacting us. The immunogen sequence for ab13604 has 99% homology with monkey. Please let me know if this is a product you wish to test and I can provide a cose for the AbTrial program.


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