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

Overview

  • Product name
    Anti-Catalase antibody [EP1929Y] - Peroxisome Marker
    See all Catalase primary antibodies
  • Description
    Rabbit monoclonal [EP1929Y] to Catalase - Peroxisome Marker
  • Tested applications
    Suitable for: WB, ICC, Flow Cyt, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

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

  • Positive control
    • WB: HeLa cell lysate. IHC-P: human brain tissue.
  • General notes

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

     

    Alternative versions available:

    Anti-Catalase antibody (Alexa Fluor® 488) [EP1929Y] - Peroxisome Marker (ab185041)

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

Applications

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.

Application Abreviews Notes
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.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      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.
    • Involvement in disease
      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.
    • Sequence similarities
      Belongs to the catalase family.
    • Post-translational
      modifications
      The N-terminus is blocked.
    • Cellular localization
      Peroxisome.
    • Information by UniProt
    • Database links
    • Alternative names
      • Cas1 antibody
      • CAT antibody
      • CATA_HUMAN antibody
      • Catalase antibody
      • Cs1 antibody
      • MGC138422 antibody
      • MGC138424 antibody
      see all

    Anti-Catalase antibody [EP1929Y] - Peroxisome Marker images



    • 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.

    References for Anti-Catalase antibody [EP1929Y] - Peroxisome Marker (ab76024)

    This product has been referenced in:
    • Ma HP Hydrogen peroxide stimulates the epithelial sodium channel through a phosphatidylinositide 3-kinase-dependent pathway. J Biol Chem 286:32444-53 (2011). WB ; Xenopus laevis . Read more (PubMed: 21795700) »
    • Kemp K  et al. Mesenchymal stem cells restore frataxin expression and increase hydrogen peroxide scavenging enzymes in friedreich ataxia fibroblasts. PLoS One 6:e26098 (2011). WB ; Human . Read more (PubMed: 22016819) »

    See all 2 Publications for this product

    Product Wall

    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
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (rat intestinal epithelial cells (RIE-1))
    Permeabilization
    Yes - 0.2% TritonX-100
    Specification
    rat intestinal epithelial cells (RIE-1)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
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    Submitted Feb 12 2016

    Application
    Flow Cytometry
    Sample
    Mouse Cell (Mouse Lewis Lung Carcinoma cells)
    Permeabilization
    Yes - 0.2% Triton X-100
    Gating Strategy
    No gating
    Specification
    Mouse Lewis Lung Carcinoma cells
    Preparation
    Cell harvesting/tissue preparation method: trypsin
    Sample buffer: Hanks’ Balanced Salt solution (HBSS)
    Fixation
    Paraformaldehyde
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    Submitted Jan 25 2016

    Application
    IHC - Wholemount
    Sample
    Human Tissue (Ovary adenocarcinoma)
    Specification
    Ovary adenocarcinoma
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    Submitted Dec 21 2015

    Application
    IHC - Wholemount
    Sample
    Mouse Tissue (Transplanted lung tumor section)
    Specification
    Transplanted lung tumor section
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    Submitted Dec 21 2015

    Application
    Western blot
    Sample
    African Green Monkey Cell lysate - whole cell (COS-7 cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris)
    Loading amount
    100000 cells
    Treatment
    30 nM Bortezomib for 24 hours
    Specification
    COS-7 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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    Submitted Jun 24 2015

    Application
    Flow Cytometry
    Sample
    Human Cell (A549 lung cancer cells)
    Permeabilization
    Yes - 0.2% Triton X-100
    Gating Strategy
    No gating
    Specification
    A549 lung cancer cells
    Preparation
    Cell harvesting/tissue preparation method: trypsin
    Sample buffer: Hanks’ Balanced Salt solution (HBSS)
    Fixation
    Paraformaldehyde
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    Submitted May 15 2015

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Lewis lung cancer, FL5.12, embryonic fibroblasts)
    Gel Running Conditions
    Reduced Denaturing (4-20% Bis-Tris)
    Loading amount
    100000 cells
    Specification
    Lewis lung cancer, FL5.12, embryonic fibroblasts
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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    Submitted May 11 2015

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (mouse embryonic fibroblasts)
    Permeabilization
    Yes - 0.2% Triton X-100
    Specification
    mouse embryonic fibroblasts
    Blocking step
    Serum as blocking agent for 15 minute(s) · Concentration: 1.5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
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    Submitted May 05 2015

    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
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    Submitted Apr 21 2015

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