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
  • 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



    • Predicted band size : 60 kDa

      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

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this ha...

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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

    Abcam user community

    Verified customer

    提交于 Apr 21 2015

    Thank you for contacting us.

    I would like to apologize for this error. When we received the information, that the antibody has been tested in the laboratory with negative results on rat tissue, we updated immediately the datasheet. This was l...

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    The ab16731 is not specific to N-terminal end of the protein. It is produced using recombinant fragment of whole protein and because it is a polyclonal antibody so it will bind to multiple epitopes. It is likely that ab16731 will be able to detect the ...

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    Thank you for sending the image which is very self explanatory for multiple bands. Please ignore my suggestions for multiple bands as these were more general rather than related to this case. The antibody (ab98210) I recommended was the second most...

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    Thank you for your email. I am sorry to hear that the results did not improve. The multiple band problem could be due to following reasons: - High protein conc. of lysates per well - High conc. of primary and secondary ab - Cell confluence or ...

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    Thank you for contacting Abcam Technical Team and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this product. I have been discussing this issue with some colleagu...

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

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