重组Anti-ADAR1抗体[EPR7033] (ab126745)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7033] to ADAR1
- Suitable for: WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-ADAR1抗体[EPR7033]
参阅全部 ADAR1 一抗 -
描述
兔单克隆抗体[EPR7033] to ADAR1 -
宿主
Rabbit -
特异性
The immunogen is designed to detect the p150 isoform and not the p110.
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经测试应用
适用于: WB, IHC-P, Flow Cyt (Intra)more details
不适用于: ICC/IF or IP -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human ADAR1 aa 200-300. The exact sequence is proprietary. The immunogen used to raise this antibody is designed to detect isoform 1 (p150) and isoforms 2-4. It does not detect Isoform 5 (p110).
Database link: P55265 -
阳性对照
- WB: HEK293T, HeLa, Ramos and SH-SY5Y cell lysates. IHC-P: Human brain tissue Flow Cyt (intra): HeLa cells
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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 +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR7033 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab126745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 |
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WB |
1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 136 kDa).
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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WB
1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 136 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
Converts multiple adenosines to inosines and creates I/U mismatched base pairs in double-helical RNA substrates without apparent sequence specificity. Has been found to modify more frequently adenosines in AU-rich regions, probably due to the relative ease of melting A/U base pairs as compared to G/C pairs. Functions to modify viral RNA genomes and may be responsible for hypermutation of certain negative-stranded viruses. Edits the messenger RNAs for glutamate receptor (GLUR) subunits by site-selective adenosine deamination. Produces low-level editing at the GLUR-B Q/R site, but edits efficiently at the R/G site and HOTSPOT1. Binds to short interfering RNAs (siRNA) without editing them and suppresses siRNA-mediated RNA interference. Binds to ILF3/NF90 and up-regulates ILF3-mediated gene expression. -
组织特异性
Ubiquitously expressed, highest levels were found in brain and lung. -
疾病相关
Defects in ADAR are a cause of dyschromatosis symmetrical hereditaria (DSH) [MIM:127400]; also known as reticulate acropigmentation of Dohi. DSH is a pigmentary genodermatosis of autosomal dominant inheritance characterized by a mixture of hyperpigmented and hypopigmented macules distributed on the dorsal parts of the hands and feet. -
序列相似性
Contains 1 A to I editase domain.
Contains 2 DRADA repeats.
Contains 3 DRBM (double-stranded RNA-binding) domains. -
翻译后修饰
Sumoylation reduces RNA-editing activity. -
细胞定位
Cytoplasm. Nucleus > nucleolus. Isoform 1 is found predominantly in cytoplasm but appears to shuttle between the cytoplasm and nucleus. Isoform 5 is found exclusively in the nucleolus. - Information by UniProt
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数据库链接
- Entrez Gene: 103 Human
- Omim: 146920 Human
- SwissProt: P55265 Human
- Unigene: 12341 Human
- Unigene: 679967 Human
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别名
- 136 kDa double-stranded RNA-binding protein antibody
- 136kDa double stranded RNA binding protein antibody
- Adar 1 antibody
see all
图片
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All lanes : Anti-ADAR1 antibody [EPR7033] (ab126745) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ADAR knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 136 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab126745 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.
ab126745 Anti-ADAR1 antibody [EPR7033] was shown to specifically react with ADAR1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266846 (knockout cell lysate ab257131) was used. Wild-type and ADAR1 knockout samples were subjected to SDS-PAGE. ab126745 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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ab126745, at 1/50 dilution, staining ADAR1 in paraffin-embedded Human brain tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of permeabilized Ramos cells, staining ADAR1 (red) with ab126745. 1x106 cells were collected and washed with blocking buffer. Cells were fixed with 2% paraformaldehyde, permeabilized with 1X FACS permeabilizing solution and blocked with blocking buffer for 30 minutes at room temperature. Cells were incubated with primary antibody (1/10) for 30 minutes at room temperature before a Fluorescently-conjugated secondary antibody or 30 min at room temperature. A rabbit IgG was used as a negative control (green).
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: ADAR1 knockout HAP1 cell lysate (20 µg)
Lane 3: HepG2 cell lysate (20 µg)
Lane 4: HeLa cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab126745 observed at 150 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab126745 was shown to recognize ADAR1 when ADAR1 knockout samples were used, along with additional cross-reactive bands. Wild-type and ADAR1 knockout samples were subjected to SDS-PAGE. ab126745 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-ADAR1 antibody [EPR7033] (ab126745) at 1/1000 dilution
Lane 1 : HeLa (treated with IFN-alpha) cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : Ramos cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 136 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (9)
ab126745 被引用在 9 文献中.
- Gromisch CM et al. Humanized anti-DEspR IgG4S228P antibody increases overall survival in a pancreatic cancer stem cell-xenograft peritoneal carcinomatosis ratnu/nu model. BMC Cancer 21:407 (2021). PubMed: 33853558
- Kung CP et al. Evaluating the therapeutic potential of ADAR1 inhibition for triple-negative breast cancer. Oncogene 40:189-202 (2021). PubMed: 33110236
- Yang H et al. LINC00667 promotes the proliferation, migration, and pathological angiogenesis in non-small cell lung cancer through stabilizing VEGFA by EIF4A3. Cell Biol Int 44:1671-1680 (2020). PubMed: 32281700
- Ma C et al. Circular RNA hsa_circ_0004872 inhibits gastric cancer progression via the miR-224/Smad4/ADAR1 successive regulatory circuit. Mol Cancer 19:157 (2020). PubMed: 33172486
- Yu J et al. ADAR1 p110 Enhances Adhesion of Tumor Cells to Extracellular Matrix in Hepatocellular Carcinoma via Up-Regulating ITGA2 Expression. Med Sci Monit 25:1469-1479 (2019). PubMed: 30798327
- Lazzari E et al. Alu-dependent RNA editing of GLI1 promotes malignant regeneration in multiple myeloma. Nat Commun 8:1922 (2017). PubMed: 29203771
- Shi L et al. Circular RNA expression is suppressed by androgen receptor (AR)-regulated adenosine deaminase that acts on RNA (ADAR1) in human hepatocellular carcinoma. Cell Death Dis 8:e3171 (2017). PubMed: 29144509
- Dou N et al. Aberrant overexpression of ADAR1 promotes gastric cancer progression by activating mTOR/p70S6K signaling. Oncotarget 7:86161-86173 (2016). PubMed: 27863387
- Arias C et al. KSHV 2.0: a comprehensive annotation of the Kaposi's sarcoma-associated herpesvirus genome using next-generation sequencing reveals novel genomic and functional features. PLoS Pathog 10:e1003847 (2014). WB ; Human . PubMed: 24453964