使用敲除细胞株进行验证RabMAb

Anti-Catalase抗体[EP1929Y] - Peroxisome Marker (ab76024)

概述

  • 产品名称
    Anti-Catalase抗体[EP1929Y] - Peroxisome Marker
    参阅全部 Catalase 一抗
  • 描述
    兔单克隆抗体[EP1929Y] to Catalase - Peroxisome Marker
  • 宿主
    Rabbit
  • 经测试应用
    适用于: WB, ICC, Flow Cyt, IHC-Pmore details
    不适用于: IP
  • 种属反应性
    与反应: Human
  • 免疫原

    A synthetic peptide corresponding to residues near the C terminus of human Catalase

  • 阳性对照
    • WB: HeLa cell lysate. IHC-P: human brain tissue.
  • 常规说明

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

     

    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

应用

Our Abpromise guarantee covers the use of ab76024 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 1/10000 - 1/20000. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).Can be blocked with Human Catalase peptide (ab225865).
ICC 1/100 - 1/250.
Flow Cyt 1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • 应用说明
    Is unsuitable for IP.
  • 靶标

    • 功能
      Occurs in almost all aerobically respiring organisms and serves to protect cells from the toxic effects of hydrogen peroxide. Promotes growth of cells including T-cells, B-cells, myeloid leukemia cells, melanoma cells, mastocytoma cells and normal and transformed fibroblast cells.
    • 疾病相关
      Defects in CAT are the cause of acatalasia (ACATLAS) [MIM:115500]; also known as acatalasemia. This disease is characterized by absence of catalase activity in red cells and is often associated with ulcerating oral lesions.
    • 序列相似性
      Belongs to the catalase family.
    • 翻译后修饰
      The N-terminus is blocked.
    • 细胞定位
      Peroxisome.
    • Information by UniProt
    • 数据库链接
    • 别名
      • Cas1 antibody
      • CAT antibody
      • CATA_HUMAN antibody
      • Catalase antibody
      • Cs1 antibody
      • MGC138422 antibody
      • MGC138424 antibody
      see all

    图片

    • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: CAT knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)

      Lanes 1 - 3: Merged signal (red and green). Green - ab76024 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab76024 was shown to specifically react with CAT when CAT knockout samples were used. Wild-type and CAT knockout samples were subjected to SDS-PAGE. Ab76024 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Overlay histogram showing HeLa cells stained with ab76024 (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 (ab76024, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Anti-Catalase antibody [EP1929Y] - Peroxisome Marker (ab76024) at 1/20000 dilution + HeLa cell lysate at 10 µg

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

      Predicted band size: 60 kDa
      Observed band size: 60 kDa

    • Immunohistochemical staining of Catalase in paraffin embedded human normal brain tissue using ab76024 at a 1/100 dilution.

    文献

    This product has been referenced in:
    • Vishnyakova PA  et al. Alterations in antioxidant system, mitochondrial biogenesis and autophagy in preeclamptic myometrium. BBA Clin 8:35-42 (2017). Read more (PubMed: 28736722) »
    • Qian K  et al. Capsaicin Suppresses Cell Proliferation, Induces Cell Cycle Arrest and ROS Production in Bladder Cancer Cells through FOXO3a-Mediated Pathways. Molecules 21:N/A (2016). Read more (PubMed: 27775662) »

    See all 6 Publications for this product

    客户评价及客户问答

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    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (A549 lung cancer cells)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris-glycine)
    Loading amount
    15 µg
    Specification
    A549 lung cancer cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Username

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    提交于 Apr 21 2015

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

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