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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ub-like Proteins
使用敲除细胞株进行验证重组RabMAb

重组Anti-USP9x抗体[EPR13809(B)] - N-terminal (ab180191)

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  • SDS
  • Certificate of Compliance
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Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
  • Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
  • Immunocytochemistry/ Immunofluorescence - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
  • Flow Cytometry (Intracellular) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
  • Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
  • Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
  • Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
  • Anti-USP9x antibody [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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 750 Alkaline Phosphatase APC Carrier Free HRP PE

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概述

  • 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.
  • 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
    For more information see here.

    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

  • 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...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    EPR13809(B)
  • 同种型

    IgG
  • 研究领域

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ub-like Proteins
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Deubiquitination
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Epigenetics and Nuclear Signaling
    • Ubiquitin & Ubiquitin Like Modifiers
    • Deubiquitination

相关产品

  • Alternative Versions

    • Anti-USP9x antibody [EPR13809(B)] - BSA and Azide free (ab250191)
    • Alexa Fluor® 568 Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab312497)
    • Alexa Fluor® 750 Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab321230)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human USP9X knockout HeLa cell line (ab265665)
  • KO cell lysates

    • Human USP9X knockout HeLa cell lysate (ab257790)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • Hep G2 whole cell lysate (ab166833)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
  • Related Products

    • Recombinant Human USP9x protein (ab152844)

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.

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)
1/1000 - 1/10000. Predicted molecular weight: 292 kDa.
ICC/IF
1/100 - 1/250.
IP
1/20.
说明
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

  • 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.
  • Target information above from: UniProt accession Q93008 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • 别名

      • Deubiquitinating enzyme FAF X antibody
      • Deubiquitinating enzyme FAF-X antibody
      • DFFRX antibody
      • Drosophila fat facets related X linked antibody
      • FAF antibody
      • Fafl antibody
      • Fam antibody
      • Fat facets homolog antibody
      • Fat facets in mammals antibody
      • Fat facets protein related X linked antibody
      • Fat facets protein related, X-linked antibody
      • Fat facets protein-related antibody
      • hFAM antibody
      • MRX99 antibody
      • Probable ubiquitin carboxyl terminal hydrolase FAF X antibody
      • Probable ubiquitin carboxyl-terminal hydrolase FAF-X antibody
      • Ubiquitin carboxyl-terminal hydrolase FAM antibody
      • Ubiquitin specific peptidase 9 X linked antibody
      • Ubiquitin specific peptidase 9, X-linked antibody
      • Ubiquitin specific processing protease FAF X antibody
      • Ubiquitin specific protease 9 X chromosome antibody
      • Ubiquitin thioesterase FAF X antibody
      • Ubiquitin thiolesterase FAF X antibody
      • Ubiquitin thiolesterase FAF-X antibody
      • Ubiquitin-specific protease 9 antibody
      • Ubiquitin-specific-processing protease FAF-X antibody
      • USP9 (gene name) antibody
      • Usp9x antibody
      • USP9X_HUMAN antibody
      • Uubiquitin specific protease 9, X chromosome (fat facets like Drosophila) antibody
      • X chromosome antibody
      • X-linked antibody
      see all

    图片

    • Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

      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.

    • Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

      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 lysate

      For 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Immunocytochemistry/ Immunofluorescence - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

      Immunofluorescent analysis of HeLa cells labeling USP9x with ab180191 at 1/100 (green) and DAPI staining (blue).

    • Flow Cytometry (Intracellular) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Flow Cytometry (Intracellular) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

      Intracellular flow cytometrical analysis of permeabilized Jurkat cells labeling USP9x with ab180191 at 1/10 (red) or a rabbit IgG negative (green). 

       

       

    • Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

      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 lysate

      For 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.

    • Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

      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.

    • Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

      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.

    • Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
      Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

    实验方案

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    文献 (6)

    发表研究结果有使用 ab180191?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab180191 被引用在 6 文献中.

    • Roldán-Romero JM  et al. Deubiquitinase USP9X loss sensitizes renal cancer cells to mTOR inhibition. Int J Cancer 153:1300-1312 (2023). PubMed: 37260183
    • Ye M  et al. TBK1 Knockdown Alleviates Axonal Transport Deficits in Retinal Ganglion Cells Via mTORC1 Activation in a Retinal Damage Mouse Model. Invest Ophthalmol Vis Sci 64:1 (2023). PubMed: 37395713
    • 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

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    Western blot abreview for Anti-USP9x antibody [EPR13809(B)] - N-terminal

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HAP1 cell line)
    Gel Running Conditions
    Non-reduced Denaturing (4-12 % Bis-Tris gel)
    Loading amount
    50 µg
    Specification
    HAP1 cell line
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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