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使用敲除细胞株进行验证重组RabMAb

重组Anti-USP10抗体[EPR4261] (ab109219)

Reviews (1) Submit a question References (8)
Western blot - Anti-USP10 antibody [EPR4261] (ab109219)
  • Western blot - Anti-USP10 antibody [EPR4261] (ab109219)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP10 antibody [EPR4261] (ab109219)
  • Immunocytochemistry/ Immunofluorescence - Anti-USP10 antibody [EPR4261] (ab109219)
  • Immunoprecipitation - Anti-USP10 antibody [EPR4261] (ab109219)
  • Anti-USP10 antibody [EPR4261] (ab109219)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4261] to USP10
  • Suitable for: WB, IP, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 750 APC Carrier Free PE

概述

  • 产品名称

    Anti-USP10抗体[EPR4261]
    参阅全部 USP10 一抗

  • 描述

    兔单克隆抗体[EPR4261] to USP10

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IP, IHC-P, ICC/IFmore details
    不适用于: Flow Cyt

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: HeLa, 293T, A375 and A549 cell lysates. IHC-P: Human colon tissue. ICC: MCF-7 IP: HeLa.

  • 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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab109219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
WB (1)
1/1000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 87 kDa).
IP
1/10 - 1/100.
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.
ICC/IF
1/250 - 1/500.
说明
WB
1/1000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 87 kDa).
IP
1/10 - 1/100.
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.
ICC/IF
1/250 - 1/500.
应用说明
Is unsuitable for Flow Cyt.

靶标

  • 功能

    Hydrolase that can remove conjugated ubiquitin from target proteins such as p53/TP53, SNX3 and CFTR. Acts as an essential regulator of p53/TP53 stability: in unstressed cells, specifically deubiquitinates p53/TP53 in the cytoplasm, leading to counteract MDM2 action and stabilize p53/TP53. Following DNA damage, translocates to the nucleus and deubiquitinates p53/TP53, leading to regulate the p53/TP53-dependent DNA damage response. Does not deubiquitinate MDM2. Deubiquitinates CFTR in early endosomes, enhancing its endocytic recycling.

  • 组织特异性

    Widely expressed.

  • 序列相似性

    Belongs to the peptidase C19 family. USP10 subfamily.

  • 翻译后修饰

    Phosphorylated by ATM following DNA damage, leading to stablization and translocation it to the nucleus.

  • 细胞定位

    Cytoplasm. Nucleus. Early endosome. Cytoplasmic in normal conditions. After DNA damage, translocates to the nucleus following phosphorylation by ATM.
  • Information by UniProt

  • 数据库链接

    • 别名

      • Deubiquitinating enzyme 10 antibody
      • KIAA0190 antibody
      • MGC124997 antibody
      • MGC2621 antibody
      • Ubiquitin carboxyl terminal hydrolase 10 antibody
      • Ubiquitin carboxyl-terminal hydrolase 10 antibody
      • Ubiquitin specific peptidase 10 antibody
      • Ubiquitin specific processing protease 10 antibody
      • Ubiquitin specific protease 10 antibody
      • Ubiquitin thioesterase 10 antibody
      • Ubiquitin thiolesterase 10 antibody
      • Ubiquitin-specific-processing protease 10 antibody
      • UBP10_HUMAN antibody
      • UBPO antibody
      • USP 10 antibody
      • USP10 antibody
      see all

    图片

    • Western blot - Anti-USP10 antibody [EPR4261] (ab109219)
      Western blot - Anti-USP10 antibody [EPR4261] (ab109219)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: USP10 knockout HAP1 cell lysate (20 µg)
      Lane 3: A549 cell lysate (20 µg)
      Lane 4: A375 cell lysate (20 µg)
      Lanes 1 to 4: Merged signal (red and green). Green - ab109219 observed at 115 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab109219 was shown to recognize USP10 when USP10 knockout samples were used, along with additional cross-reactive bands. Wild-type and USP10 knockout samples were subjected to SDS-PAGE. ab109219 and ab8245 (loading control to GAPDH) were both diluted at 1/1000 and 1/10000 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 1/10000 dilution for 1 h at room temperature before imaging.

    • Western blot - Anti-USP10 antibody [EPR4261] (ab109219)
      Western blot - Anti-USP10 antibody [EPR4261] (ab109219)
      All lanes : Anti-USP10 antibody [EPR4261] (ab109219) at 1/1000 dilution

      Lane 1 : Hela cell lysate
      Lane 2 : 293T cell lysate
      Lane 3 : A375 cell lysate
      Lane 4 : A549 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 87 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP10 antibody [EPR4261] (ab109219)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP10 antibody [EPR4261] (ab109219)

      ab109219, at a 1/250 dilution, staining USP10 in paraffin embedded Human colon tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-USP10 antibody [EPR4261] (ab109219)
      Immunocytochemistry/ Immunofluorescence - Anti-USP10 antibody [EPR4261] (ab109219)

      ICC/IF image of ab109219 stained MCF-7 cells. The cells were 100% methanol fixed (5 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 ab109219 at 1/50 dilution overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

    • Immunoprecipitation - Anti-USP10 antibody [EPR4261] (ab109219)
      Immunoprecipitation - Anti-USP10 antibody [EPR4261] (ab109219)

      USP10 was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg with 109219 at 1/30 dilution (2µg) . VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg

      Lane 2: ab109219 IP in HeLa whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab109219 in HeLa whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Fresh lysate should be used to minimize protein degradation.

    • Anti-USP10 antibody [EPR4261] (ab109219)
      Anti-USP10 antibody [EPR4261] (ab109219)

    文献 (8)

    发表研究结果有使用 ab109219？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab109219 被引用在 8 文献中.

    • Xin SL  et al. USP10 Alleviates Palmitic Acid-induced Steatosis through Autophagy in HepG2 Cells. J Clin Transl Hepatol 11:45-57 (2023). PubMed: 36406315
    • Cao YF  et al. Targeting USP10 induces degradation of oncogenic ANLN in esophageal squamous cell carcinoma. Cell Death Differ 30:527-543 (2023). PubMed: 36526897
    • Qiu W  et al. USP10 deubiquitinates RUNX1 and promotes proneural-to-mesenchymal transition in glioblastoma. Cell Death Dis 14:207 (2023). PubMed: 36949071
    • Nakken S  et al. AGAP2-AS1 as a prognostic biomarker in low-risk clear cell renal cell carcinoma patients with progressing disease. Cancer Cell Int 21:690 (2021). PubMed: 34930263
    • Zeng Z  et al. Association and clinical implication of the USP10 and MSH2 proteins in non-small cell lung cancer. Oncol Lett 17:1128-1138 (2019). PubMed: 30655874
    • Peng XC  et al. Clinical significance of TM4SF1 as a tumor suppressor gene in gastric cancer. Cancer Med 7:2592-2600 (2018). PubMed: 29665316
    • Chen Q  et al. USP10 promotes proliferation and migration and inhibits apoptosis of endometrial stromal cells in endometriosis through activating the Raf-1/MEK/ERK pathway. Am J Physiol Cell Physiol 315:C863-C872 (2018). PubMed: 30281322
    • Zeng Z  et al. Myxoid adrenocortical adenoma with a pseudoglandular pattern: a case report and literature review. Int J Clin Exp Pathol 10:8908-8915 (2017). PubMed: 31966759

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    Western blot abreview for Anti-USP10 antibody [EPR4261]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (EC109)
    Gel Running Conditions
    Non-reduced Denaturing (10%SDS)
    Loading amount
    80 µg
    Specification
    EC109
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°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.

    Qingyuan Yang

    Verified customer

    提交于 Jan 08 2016

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