重组Anti-Dnmt1抗体[EPR3522] (ab92314)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3522] to Dnmt1
- Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Dnmt1抗体[EPR3522]
参阅全部 Dnmt1 一抗 -
描述
兔单克隆抗体[EPR3522] to Dnmt1 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IP, ICC/IFmore details
不适用于: ChIP or IHC-P -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human Dnmt1 aa 1600-1700 (C terminal). The exact sequence is proprietary.
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阳性对照
- WB: HuT-78, Jurkat or 293T lysate. ICC: Jurkat cells. Flow Cyt: HeLa cells IP: Hut-78.
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常规说明
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR3522 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab92314于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
Flow Cyt (Intra) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB |
1/1000 - 1/10000. Predicted molecular weight: 183 kDa.
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IP |
1/20.
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ICC/IF |
1/50 - 1/100.
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说明 |
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Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Predicted molecular weight: 183 kDa. |
IP
1/20. |
ICC/IF
1/50 - 1/100. |
靶标
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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
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数据库链接
- Entrez Gene: 1786 Human
- Omim: 126375 Human
- SwissProt: P26358 Human
- Unigene: 202672 Human
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别名
- ADCADN antibody
- AIM antibody
- CXXC finger protein 9 antibody
see all
图片
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Dnmt1 was immunoprecipitated from 0.35 mg Hut-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate 10 µg with 92314 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Hut-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate 10 µg
Lane 2: ab92314 IP in Hut-78 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab92314 in Hut-78 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: DNMT1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HEK293 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab92314 observed at 170 kDa. Red - loading control, ab18058, observed at 130 kDa.ab92314 was shown to specifically react with DNMT1 when DNMT1 knockout samples were used. Wild-type and DNMT1 knockout samples were subjected to SDS-PAGE. Ab92314 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescence staining of Jurkat cells with purified ab92314 at a working dilution of 1/2000, counter-stained with DAPI. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-rabbit (ab150077), used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.
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Overlay histogram showing HeLa cells stained with ab92314 (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 followed by the antibody (ab92314, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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All lanes : Anti-Dnmt1 antibody [EPR3522] (ab92314) at 1/1000 dilution
Lane 1 : HuT-78 cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : 293T cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 183 kDa
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (12)
ab92314 被引用在 12 文献中.
- Huang K & Tang Y SChLAP1 promotes prostate cancer development through interacting with EZH2 to mediate promoter methylation modification of multiple miRNAs of chromosome 5 with a DNMT3a-feedback loop. Cell Death Dis 12:188 (2021). PubMed: 33589600
- Reilly BM et al. 5-Azacytidine Transiently Restores Dysregulated Erythroid Differentiation Gene Expression in TET2-Deficient Erythroleukemia Cells. Mol Cancer Res 19:451-464 (2021). PubMed: 33172974
- Du Q et al. DNA methylation is required to maintain both DNA replication timing precision and 3D genome organization integrity. Cell Rep 36:109722 (2021). PubMed: 34551299
- Yang E et al. Epigenetic silencing of UBXN8 contributes to leukemogenesis in t(8;21) acute myeloid leukemia. Exp Mol Med 53:1902-1910 (2021). PubMed: 34921223
- Recagni M et al. The Role of Alternative Lengthening of Telomeres Mechanism in Cancer: Translational and Therapeutic Implications. Cancers (Basel) 12:N/A (2020). PubMed: 32290440