重组Anti-USP9x抗体[EPR13809(B)] - N-terminal (ab180191)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR13809(B)] to USP9x - N-terminal
- Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-USP9x抗体[EPR13809(B)] - N-terminal
参阅全部 USP9x 一抗 -
描述
兔单克隆抗体[EPR13809(B)] to USP9x - N-terminal -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), IHC-P, WB, ICC/IF, IPmore details -
种属反应性
与反应: Mouse, Human
预测可用于: Rat -
免疫原
Synthetic peptide within Human USP9x aa 1-100 (Cysteine residue). The exact sequence is proprietary.
Database link: Q93008 -
阳性对照
- WB: HeLa, Hap1, T84, NIH3T3, Jurkat and HepG2 cell lysates. IHC-P: Human kidney and testis tissues. IP: HeLa and NIH3T3 cell lysates. Flow Cyt (intra): Jurkat cells. ICC/IF: 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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 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
EPR13809(B) -
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
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Positive Controls
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab180191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) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Antigen retrieval is recommended. |
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 292 kDa.
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ICC/IF |
1/100 - 1/250.
|
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IP |
1/20.
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说明 |
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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. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Antigen retrieval is recommended. |
WB
1/1000 - 1/10000. Predicted molecular weight: 292 kDa. |
ICC/IF
1/100 - 1/250. |
IP
1/20. |
靶标
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功能
Deubiquitinase involved both in the processing of ubiquitin precursors and of ubiquitinated proteins. May therefore play an important role regulatory role at the level of protein turnover by preventing degradation of proteins through the removal of conjugated ubiquitin. Essential component of TGF-beta/BMP signaling cascade. Regulates chromosome alignment and segregation in mitosis by regulating the localization of BIRC5/survivin to mitotic centromeres. Specifically hydrolyzes both 'Lys-29'- and 'Lys-33'-linked polyubiquitins chains. Specifically deubiquitinates monoubiquitinated SMAD4, opposing the activity of E3 ubiquitin-protein ligase TRIM33. -
组织特异性
Widely expressed in embryonic and adult tissues. -
序列相似性
Belongs to the peptidase C19 family. -
细胞定位
Cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 8239 Human
- Entrez Gene: 22284 Mouse
- Entrez Gene: 363445 Rat
- Omim: 300072 Human
- SwissProt: Q93008 Human
- SwissProt: P70398 Mouse
- Unigene: 77578 Human
- Unigene: 242646 Mouse
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别名
- Deubiquitinating enzyme FAF X antibody
- Deubiquitinating enzyme FAF-X antibody
- DFFRX antibody
see all
图片
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All lanes : Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : USP9X knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 292 kDa
Observed band size: 290 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab180191 observed at 290 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab180191 was shown to react with USP9x in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265665 (knockout cell lysate ab257790) was used. Wild-type HeLa and USP9X knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab180191 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 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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Immunohistochemical analysis of paraffin embedded Human kidney tissue labeling USP9x with ab180191 at 1/50.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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ab180191 (purified) at 1/20 dilution immunoprecipitating ETS2 in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg.
Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+): ab180191 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab180191 in HeLa whole cell lysateFor western blotting, ab180191 at 1/500 dilution (0.23 µg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.
Blocking and diluting buffer: 5% NFDM /TBST.
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Immunofluorescent analysis of HeLa cells labeling USP9x with ab180191 at 1/100 (green) and DAPI staining (blue).
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Intracellular flow cytometrical analysis of permeabilized Jurkat cells labeling USP9x with ab180191 at 1/10 (red) or a rabbit IgG negative (green).
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ab180191 (purified) at 1/20 dilution immunoprecipitating ETS2 in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 µg.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 µg.
Lane 2 (+): ab180191 & NIH/3T3 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab180191 in NIH/3T3 whole cell lysateFor western blotting, ab180191 at 1/500 dilution (0.23 µg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.
Blocking and diluting buffer: 5% NFDM /TBST.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: USP9x knockout HAP1 cell lysate (20 µg)
Lane 3: T84 cell lysate (20 µg)
Lane 4: NIH3T3 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab180191 observed at 270 kDa. Red - loading control, ab18058, observed at 117 kDa.ab180191 was shown to specifically react with USP9x when USP9x knockout samples were used. Wild-type and USP9x knockout samples were subjected to SDS-PAGE. ab180191 and ab18058 (loading control to Vinculin) were diluted at 1 µg/ml 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.
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All lanes : Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 292 kDa -
Immunohistochemical analysis of paraffin embedded Human testis tissue labeling USP9x with ab180191 at 1/50.
Perform heat mediated antigen retrieval 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 (4)
ab180191 被引用在 4 文献中.
- Du M et al. Single-cell RNA sequencing reveals that BMPR2 mutation regulates right ventricular function via ID genes. Eur Respir J 60:N/A (2022). PubMed: 34857612
- Chen W et al. USP9X promotes apoptosis in cholangiocarcinoma by modulation expression of KIF1Bß via deubiquitinating EGLN3. J Biomed Sci 28:44 (2021). PubMed: 34112167
- Chen MY et al. USP9X promotes the progression of hepatocellular carcinoma by regulating beta-catenin. Ir J Med Sci 189:865-871 (2020). PubMed: 32065347
- Li H et al. VCP/p97 increases BMP signaling by accelerating ubiquitin ligase Smurf1 degradation. FASEB J 33:2928-2943 (2019). PubMed: 30335548