重组Anti-RNA Helicase A抗体[EPR13521] (ab183731)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR13521] to RNA Helicase A
- Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-RNA Helicase A抗体[EPR13521]
参阅全部 RNA Helicase A 一抗 -
描述
兔单克隆抗体[EPR13521] to RNA Helicase A -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), IHC-P, WB, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, 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, MCF7 and THP-1 cell lysates; Mouse and rat brain, liver and testis tissue lysates. IHC-P: Human kidney carcinoma and lung adenocarcinoma tissue, Human and mouse breast carcinoma, human, mouse and rat testis tissue. ICC/IF: HeLa cells 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.
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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR13521 -
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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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab183731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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Flow Cyt (Intra) |
1/10 - 1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P |
1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/50 - 1/100. |
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WB |
1/10000 - 1/50000. Detects a band of approximately 141 kDa (predicted molecular weight: 141 kDa).
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ICC/IF |
1/50.
For unpurified use at 1/250. |
说明 |
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Flow Cyt (Intra)
1/10 - 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IHC-P
1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/50 - 1/100. |
WB
1/10000 - 1/50000. Detects a band of approximately 141 kDa (predicted molecular weight: 141 kDa). |
ICC/IF
1/50. For unpurified use at 1/250. |
靶标
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功能
Unwinds double-stranded DNA and RNA in a 3' to 5' direction. Alteration of secondary structure may subsequently influence interactions with proteins or other nucleic acids. Functions as a transcriptional activator. Component of the CRD-mediated complex that promotes MYC mRNA stability. -
序列相似性
Belongs to the DEAD box helicase family. DEAH subfamily.
Contains 2 DRBM (double-stranded RNA-binding) domains.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain. -
结构域
The MTAD domain mediates interaction with the RNA polymerase II holoenzyme. The NTD domain is necessary and sufficient for nucleo-cytoplasmic shuttling and interaction with HRMT1L2 and SMN1. -
翻译后修饰
Methylated. HRMT1L2 mediated methylation of undefined Arg residues in the NTD is required for nuclear localization.
May be phosphorylated by PRKDC/XRCC7. Phosphorylated upon DNA damage, probably by ATM or ATR. -
细胞定位
Nucleus > nucleolus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Can shuttle between nucleus and cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 1660 Human
- Entrez Gene: 13211 Mouse
- Entrez Gene: 304859 Rat
- Omim: 603115 Human
- SwissProt: Q08211 Human
- SwissProt: O70133 Mouse
- Unigene: 191518 Human
- Unigene: 20000 Mouse
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别名
- ATP dependent RNA helicase A antibody
- ATP-dependent RNA helicase A antibody
- DDX 9 antibody
see all
图片
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Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.Nuclear staining on rat testis. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.Nuclear staining on mouse testis. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.Nuclear staining on human testis. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemical analysis of paraffin-embedded Mouse breast carcinoma tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.Nuclear staining on mouse breast carcinoma. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.Nuclear staining on human breast carcinoma. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
All lanes : Anti-RNA Helicase A antibody [EPR13521] (ab183731) at 1/1000 dilution
Lane 1 : Rat brain tissue lysate
Lane 2 : Rat liver tissue lysate
Lane 3 : Rat testis tissue lysate
Lane 4 : Mouse brain tissue lysate
Lane 5 : Mouse liver tissue lysate
Lane 6 : Mouse testis tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 141 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as a GAPDH loading control.
Exposure time: Lanes 1-5: 103 seconds, Lane 6: 26 seconds.
Lysates were freshly made and used immediately to minimize protein degradation.
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All lanes : Anti-RNA Helicase A antibody [EPR13521] (ab183731) at 1/10000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 3 : THP-1 (Human monocytic leukemia monocyte) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 141 kDa
Observed band size: 128,141 kDa why is the actual band size different from the predicted?Blocking/Diluting Buffer and concentration: 5% NFDM/TBST
The observed bands are consistent with what are described in PMID: 28337295
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Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RNA Helicase A with purified ab183731 at 1/20 dilution (5µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RNA Helicase A with purified ab183731 at 1/50 dilution (2.2 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney carcinoma tissue sections labeling RNA Helicase A with purified ab183731 at 1/400 dilution (0.28 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Intracellular Flow Cytometry analysis of HeLa cells using ab183731 (unpurified) at a 1/10 dilution (red) or a Rabbit monoclonal IgG (negative) (green). Goat anti rabbit IgG (FITC) secondary used at a 1/150 dilution.
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Immunocytochemistry analysis of HeLa cells (fixative -20℃ Acetone) labeling RNA Helicase A with ab183731 (unpurified) at a 1/250 dilution. Goat anti rabbit IgG (Alexa Fluor® 488) secondary used at a 1/200 diution.
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Immunohistochemical analysis of paraffin embedded Human lung adenocarcinoma tissue labeling RNA Helicase A with ab183731 (unpurified) at a 1/100 dilution. Prediluted HRP conjugated Rabbit IgG secondary used. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with EDTA buffer pH 9 before commencing with IHC staining protocol.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (6)
ab183731 被引用在 6 文献中.
- Xiao L et al. LncRNA PCAT19 induced by SP1 and acted as oncogene in gastric cancer competitively binding to miR429 and upregulating DHX9. J Cancer 13:102-111 (2022). PubMed: 34976174
- Alagia A et al. Proximity Ligation Assay for Detection of R-Loop Complexes upon DNA Damage. Methods Mol Biol 2528:289-303 (2022). PubMed: 35704199
- Lin YC et al. Oxaliplatin-Induced DHX9 Phosphorylation Promotes Oncogenic Circular RNA CCDC66 Expression and Development of Chemoresistance. Cancers (Basel) 12:N/A (2020). PubMed: 32187976
- Ilik IA et al. SON and SRRM2 are essential for nuclear speckle formation. Elife 9:N/A (2020). PubMed: 33095160
- Panhale A et al. CAPRI enables comparison of evolutionarily conserved RNA interacting regions. Nat Commun 10:2682 (2019). PubMed: 31213602
- Aktas T et al. DHX9 suppresses RNA processing defects originating from the Alu invasion of the human genome. Nature 544:115-119 (2017). PubMed: 28355180