製品の概要
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製品名Anti-Exportin-5 antibody
Exportin-5 一次抗体 製品一覧 -
製品の詳細Mouse monoclonal to Exportin-5
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由来種Mouse
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アプリケーション適用あり: WB, Flow Cyt, IP, ICC/IF, IHC-Pmore details
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種交差性交差種: Human
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免疫原
Recombinant full length protein, corresponding to amino acids 1-281 of Human Exportin-5
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特記事項
Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab129006 as a replacement.
製品の特性
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製品の状態Liquid
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保存方法Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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バッファーPreservative: None
PBS, pH 7.2 -
Concentration information loading... -
精製度Protein G purified
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ポリ/モノモノクローナル
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アイソタイプIgG1
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軽鎖の種類kappa
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研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Related Products
アプリケーション
Our Abpromise guarantee covers the use of ab57491 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| WB | Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 136 kDa. | |
| Flow Cyt | Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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| IP | Use at an assay dependent concentration. | |
| ICC/IF | Use a concentration of 10 µg/ml. | |
| IHC-P | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ターゲット情報
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機能Mediates the nuclear export of proteins bearing a double-stranded RNA binding domain (dsRBD) and double-stranded RNAs (cargos). XPO5 in the nucleus binds cooperatively to the RNA and to the GTPase Ran in its active GTP-bound form. Proteins containing dsRBDs can associate with this trimeric complex through the RNA. Docking of this complex to the nuclear pore complex (NPC) is mediated through binding to nucleoporins. Upon transit of a nuclear export complex into the cytoplasm, hydrolysis of Ran-GTP to Ran-GDP (induced by RANBP1 and RANGAP1, respectively) cause disassembly of the complex and release of the cargo from the export receptor. XPO5 then returns to the nuclear compartment by diffusion through the nuclear pore complex, to mediate another round of transport. The directionality of nuclear export is thought to be conferred by an asymmetric distribution of the GTP-and GDP-bound forms of Ran between the cytoplasm and nucleus. Overexpression may in some circumstances enhance RNA-mediated gene silencing (RNAi).
Mediates the nuclear export of micro-RNA precursors, which form short hairpins. Also mediates the nuclear export of synthetic short hairpin RNAs used for RNA interference, and adenovirus VA1 dsRNA. In some circumstances can also mediate the nuclear export of deacylated and aminoacylated tRNAs. Specifically recognizes dsRNAs that lack a 5'-overhang in a sequence-independent manner, have only a short 3'-overhang, and that have a double-stranded length of at least 15 base-pairs. Binding is dependent on Ran-GTP. -
組織特異性Expressed in heart, brain, placenta, lung, skeletal muscle, kidney and pancreas.
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配列類似性Belongs to the exportin family.
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細胞内局在Nucleus. Cytoplasm. Shuttles between the nucleus and the cytoplasm.
- Information by UniProt
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参照データベース
- Entrez Gene: 57510 Human
- Omim: 607845 Human
- SwissProt: Q9HAV4 Human
- Unigene: 203206 Human
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別名
- Exp 5 antibody
- Exp5 antibody
- Exportin 5 antibody
see all
画像
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Western blot - Anti-Exportin-5 antibody (ab57491)Exportin-5 antibody (ab57491) at 1ug/lane + HeLa cell lysate at 25ug/lane. -
Immunoprecipitation - Anti-Exportin-5 antibody (ab57491)Exportin-5 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 10µg of Mouse monoclonal to Exportin-5 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab57491.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 140kDa: Exportin-5; non specific - 65kDa: We are unsure as to the identity of this extra band. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Exportin-5 antibody (ab57491)IHC image of ab57491 staining in normal human testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab57491, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Immunocytochemistry/ Immunofluorescence - Anti-Exportin-5 antibody (ab57491)ICC/IF image of ab57491 stained HeLa cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab57491, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879 Dylight 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. -
Flow Cytometry - Anti-Exportin-5 antibody (ab57491)Overlay histogram showing HEK293 cells stained with ab57491 (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 (ab57491, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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.
プロトコール
参考文献
This product has been referenced in:
- Guo J et al. Unraveling molecular effects of ADAR1 overexpression in HEK293T cells by label-free quantitative proteomics. Cell Cycle 15:1591-601 (2016). Read more (PubMed: 27104882) »
- Schertzer M et al. Human regulator of telomere elongation helicase 1 (RTEL1) is required for the nuclear and cytoplasmic trafficking of pre-U2 RNA. Nucleic Acids Res 43:1834-47 (2015). Human . Read more (PubMed: 25628358) »
レビューと Q&A
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"