Anti-Dnmt1抗体[60B1220.1] - ChIP Grade (ab13537)

概述

  • 产品名称Anti-Dnmt1抗体[60B1220.1] - ChIP Grade
    参阅全部 Dnmt1 一抗
  • 描述
    小鼠单克隆抗体[60B1220.1] to Dnmt1 - ChIP Grade
  • 特异性This antibody detects a ~180 kDa protein, corresponding to the apparent molecular mass of Dnmt1 on SDS-PAGE immunoblots in samples of human and mouse origin. Immunogen itself has been shown to be toxic.
  • 经测试应用适用于: IHC-P, IHC-Fr, ChIP, Flow Cyt, WB, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Zebrafish
    预测可用于: Cow
  • 免疫原

    Synthetic peptide:

    EKDDREDKENAFKR

    , corresponding to amino acids 637-650 of Human Dnmt1

  • 阳性对照
    • Human kidney (IHC), mouse ES or NIH 3T3 cell lysate (WB)
  • 常规说明For maximum product recovery, centrifuge the product vial before removing cap. Shipped at on gel packs.

性能

应用

Our Abpromise guarantee covers the use of ab13537 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
IHC-P Use a concentration of 1 - 2 µg/ml. DNMT1 high levels are toxic, as a result it may be tough to find sometimes. Signal amplification might be needed.
IHC-Fr Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.
Flow Cyt 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 183 kDa).
IP Use 2µg for 106 cells.

靶标

  • 功能Methylates CpG residues. Preferentially methylates hemimethylated DNA. Associates with DNA replication sites in S phase maintaining the methylation pattern in the newly synthesized strand, that is essential for epigenetic inheritance. Associates with chromatin during G2 and M phases to maintain DNA methylation independently of replication. It is responsible for maintaining methylation patterns established in development. DNA methylation is coordinated with methylation of histones. Mediates transcriptional repression by direct binding to HDAC2. In association with DNMT3B 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.
  • 组织特异性Ubiquitous; highly expressed in fetal tissues, heart, kidney, placenta, peripheral blood mononuclear cells, and expressed at lower levels in spleen, lung, brain, small intestine, colon, liver, and skeletal muscle. Isoform 2 is less expressed than isoform 1.
  • 序列相似性Belongs to the C5-methyltransferase family.
    Contains 2 BAH domains.
    Contains 1 CXXC-type zinc finger.
  • 结构域The N-terminal part is required for homodimerization and acts as a regulatory domain.
  • 翻译后修饰Sumoylated; sumoylation increases activity.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ADCADN antibody
    • AIM antibody
    • CXXC finger protein 9 antibody
    • CXXC-type zinc finger protein 9 antibody
    • CXXC9 antibody
    • DNA (cytosine 5 ) methyltransferase 1 antibody
    • DNA (cytosine-5)-methyltransferase 1 antibody
    • DNA methyltransferase 1 antibody
    • DNA methyltransferase HsaI antibody
    • DNA methyltransferase M.HsaI. antibody
    • DNA MTase antibody
    • DNA MTase HsaI antibody
    • DNMT 1 antibody
    • DNMT antibody
    • Dnmt1 antibody
    • DNMT1_HUMAN antibody
    • Dnmt1o antibody
    • FLJ16293 antibody
    • HSN1E antibody
    • M.HsaI antibody
    • MCMT antibody
    • Met1 antibody
    • MGC104992 antibody
    • mMmul antibody
    • MommeD2 antibody
    see all

Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade 图像

  • Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537) at 1/1000 dilution + 0.1ug mouse recombinant protein. Additional lower band is probably degraded or in other ways modified Dnmt1.

    Predicted band size : 183 kDa
  • ab13537 at 1/10000 dilution staining Dnmt1 in mouse brain tissue section by Immunohistochemistry (Bouin's fixative fixed paraffin-embedded tissue sections). Tissue underwent heat mediated antigen retrieval in microwave with two, 5 minutes incubation intervals in citrate buffer. An antibody amplifier™ was used for staining. A HRP-conjugated anti-mouse secondary was used at 1/10 dilution.

  • Overlay histogram showing HeLa cells stained with ab13537 (red line). The cells were fixed with 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 followed by the antibody (ab13537, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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 slightly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • All lanes : Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537) at 1 µg/ml

    Lane 1 : HCT116 whole cell lysate
    Lane 2 : HCT116 DNMT1 KO whole cell lysate
    Lane 3 : HCT116 DNMT3b KO whole cell lysate

    Secondary
    HRP conjugated Sheep anti-mouse IgG
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 183 kDa


    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview

    See Abreview

Anti-Dnmt1 antibody [60B1220.1] - ChIP Grade (ab13537)参考文献

This product has been referenced in:
  • Chalertpet K  et al. Human papillomavirus type 16 E7 oncoprotein mediates CCNA1 promoter methylation. Cancer Sci 106:1333-40 (2015). WB, ChIP ; Human . Read more (PubMed: 26250467) »
  • Cacan E  et al. Inhibition of HDAC1 and DNMT1 modulate RGS10 expression and decrease ovarian cancer chemoresistance. PLoS One 9:e87455 (2014). Human . Read more (PubMed: 24475290) »

See all 29 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Ferret Tissue lysate - whole (organotypic slices of whole brain tissue)
Gel Running Conditions Reduced Denaturing (3-8% Tris-acetate gel% Tris-acetate gel)
Loading amount 40 µg
Specification organotypic slices of whole brain tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Francis Djankpa

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提交于 Apr 29 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Fibroblast)
Specification Fibroblast
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
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提交于 Sep 30 2015

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Brain)
Permeabilization Yes - Triton x-100, 0.01%
Specification Brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
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提交于 Jun 17 2015

Application ChIP
Sample Mouse Cell lysate - whole cell (mouse)
Negative control IgG mouse antibody
Specification mouse
Detection step Real-time PCR
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1%formaldehyde
Positive control none
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提交于 Apr 16 2015

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (Cardiomyocytes)
Specification Cardiomyocytes
Treatment Glucose, 100mM
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Jan 15 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (4-20% gradient gel)
Sample Human Cell lysate - whole cell (HCT116)
Specification HCT116
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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提交于 Nov 03 2014

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 30 µg
Gel Running Conditions Reduced Denaturing (6%)
Sample Mouse Cell lysate - whole cell (Mouse N2A neuroblastoma)
Specification Mouse N2A neuroblastoma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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提交于 Apr 03 2014

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (6%)
Sample Human Cell lysate - whole cell (hek 293)
Specification hek 293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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提交于 Apr 02 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 23 2013

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