重组Anti-ADAR1抗体[EPR25431-60] - BSA and Azide free (ab307586)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25431-60] to ADAR1 - BSA and Azide free
- Suitable for: WB, ICC/IF, Flow Cyt (Intra), IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-ADAR1抗体[EPR25431-60] - BSA and Azide free
参阅全部 ADAR1 一抗 -
描述
兔单克隆抗体[EPR25431-60] to ADAR1 - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, Flow Cyt (Intra), IPmore details
不适用于: IHC-P -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, 293T, SH-SY5Y, HepG2, Wild-type HEK-293T, Ramos, HeLa treated with 10ng/ml IFN alpha 1 for 16 hours and Human kidney whole cell lysates. ICC: HeLa cells treated with IFN alpha 1 (human) (10 ng/ml) for 16 hours and HEK293T treated with IFN alpha 1 (10 ng/ml) for 16 hours Flow Cyt: Wild-type 293T cell. IP: HeLa cell.
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常规说明
ab307586 is the carrier-free version of ab307585
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. -
存储溶液
pH: 7.20
Constituent: 100% PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR25431-60 -
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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KO cell lines
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab307586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 |
Use at an assay dependent concentration. Predicted molecular weight: 150 kDa.
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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说明 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 150 kDa. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
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 [EPR25431-60] - BSA and Azide free (ab307586) at 1/1000 dilution
Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 20 µg
Lane 2 : 293T (human embryonic kidney epithelial cell) whole cell lysate 20 µg
Lane 3 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 20 µg
Lane 4 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 150 kDa
Observed band size: 150 kDa
Exposure time: 70 secondsThis data was developed using ab307585, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
Exposure time: 70 seconds
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All lanes : Anti-ADAR1 antibody [EPR25431-60] (ab307585) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate 20 µg
Lane 2 : ADAR1 knockout HEK-293T whole cell lysate 20 µg
Lane 3 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 20 µg
Lane 4 : Ramos (human burkitt's lymphoma b lymphocyte) whole cell lysate 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 150 kDa
Observed band size: 150 kDa
Exposure time: 59 secondsThis data was developed using ab307585, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Performed under reducing conditions.
In Western blot, ab307585 was shown to bind specifically to ADAR1. A band was observed at 150 kDa in wild-type HEK-293T cell lysates whereas no signal observed at this size in ADAR1 knockout cell line ab266846 (knockout cell lysate ab257131).Exposure time: 59 seconds
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All lanes : Anti-ADAR1 antibody [EPR25431-60] (ab307585) at 1/1000 dilution
Lane 1 : Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 20 µg
Lane 2 : HeLa treated with 10ng/ml IFN alpha 1 for 16 hours whole cell lysate 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 150 kDa
Observed band size: 150 kDa
Exposure time: 37 secondsThis data was developed using ab307585, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 37 seconds
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Human kidney tissue lysate 20 µg
Secondary
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 150 kDa
Observed band size: 150 kDa
Exposure time: 158 secondsThis data was developed using ab307585, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 158 seconds
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This data was developed using ab307585, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling ADAR1 with ab307585 at 1/500 (1.08 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing increased nuclear and cytoplasmic staining in HeLa cells treated with IFN alpha 1 (human) (10 ng/ml) for 16 hours. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
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This data was developed using ab307585, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized ADAR KO HEK293T (ab266846) cells labelling ADAR1 with ab307585 at 1/500 (1.08 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing increased nuclear and cytoplasmic staining in parental HEK293T cells treated with IFN alpha 1 (human) (10 ng/ml) for 16 hours, and no staining in treated ADAR KO HEK293T cells. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
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This data was developed using ab307585, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Wild-type 293T (human embryonic kidney epithelial cell, Right) / ADAR1 knockout 293T (Left) cells labelling ADAR1 with ab307585 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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This data was developed using ab307585, the same antibody clone in a different buffer formulation.
ADAR1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab307585 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307585 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2: abab307585 IP in HeLa whole cell lysate
Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab307585 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: Lanes 1-3: 5 seconds (left), Lanes 1-3: 3 seconds (right).
The IP experiment was performed by ab307585 using HeLa cells. On the left the IP blot was probed with ab307585 and on the right the blot was probed by another anti-ADAR1 antibody (ab126745)(1:1000 dilution).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (0)
ab307586 尚未被引用在任何文献中。