使用敲除细胞株进行验证RabMAb

Anti-USP10抗体[EPR4261] (ab109219)

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
  • 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

    A synthetic peptide corresponding to residues in Human USP10.

  • 阳性对照
    • WB: HeLa, 293T, A375 and A549 cell lysates IHC-P: Human colon tissue This antibody gave a positive result when used in the following methanol fixed cell lines: MCF-7
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    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

Our Abpromise guarantee covers the use of ab109219 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 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. 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

    Anti-USP10 antibody [EPR4261] 图像



    • Predicted band size : 87 kDa

      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 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

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

    Anti-USP10 antibody [EPR4261] (ab109219)参考文献

    ab109219 has not yet been referenced specifically in any publications.

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    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
    Username

    Miss. Qingyuan Yang

    Verified customer

    提交于 Jan 08 2016

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"