Overview

  • Product name
  • Description
    Rabbit polyclonal to Catalase
  • Host species
    Rabbit
  • Specificity
    This antibody reacts with Catalase
  • Tested applications
    Suitable for: WB, IP, ICC, IHC-Pmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues near the N terminus of human Catalase

  • Positive control
    • IHC-P: Human liver tissue WB: TF1 lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab52477 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 1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).
IP 1/10.
ICC 1/50 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt.
  • 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

    Images

    • Anti-Catalase antibody (ab52477) at 1/1000 dilution + TF1 lysate at 10 µg

      Secondary
      Goat anti-rabbit HRP labeled at 1/2000 dilution

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

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Catalase knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Jurkat cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab52477 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab52477 was shown to specifically react with Catalase when Catalase knockout samples were used. Wild-type and Catalase knockout samples were subjected to SDS-PAGE. ab52477 and ab8245 (loading control to GAPDH) were 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 (IRDye® 800CW) preadsorbed (ab216773) andGoat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • IHC image of Catalasestaining in a formalin fixed, paraffin embedded normal human liver tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab52477, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

       

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • All lanes : Anti-Catalase antibody (ab52477) at 1 µg/ml

      Lane 1 : Whole cell lysate prepared from 293T cells
      Lane 2 : Whole cell lysate prepared from HeLa cells
      Lane 3 : Whole cell lysate prepared from U251 cells
      Lane 4 : Whole cell lysate prepared from Jurkat cells
      Lane 5 : Whole cell lysate prepared from U2OS cells

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP conjugated pig anti-rabbit polyclonal at 1/5000 dilution

      Developed using the ECL technique.

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


      Exposure time: 5 minutes

      See Abreview

    • All lanes : Anti-Catalase antibody (ab52477) at 1 µg/ml

      Lane 1 : Whole cell lysate prepared from 9L cells
      Lane 2 : Whole cell lysate prepared from PC12 cells
      Lane 3 : Whole cell lysate prepared from RIE-1 cells

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP conjugated pig anti-rabbit polyclonal at 1/5000 dilution

      Developed using the ECL technique.

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


      Exposure time: 5 minutes

      See Abreview

    References

    This product has been referenced in:
    • Krishnan N  et al. Harnessing insulin- and leptin-induced oxidation of PTP1B for therapeutic development. Nat Commun 9:283 (2018). Read more (PubMed: 29348454) »
    • Fazakerley DJ  et al. Mitochondrial CoQ deficiency is a common driver of mitochondrial oxidants and insulin resistance. Elife 7:N/A (2018). Read more (PubMed: 29402381) »
    See all 17 Publications for this product

    Customer reviews and Q&As

    1-10 of 10 Abreviews or Q&A

    Application
    Western blot
    Sample
    Grey seal Tissue lysate - whole (Right ventricle, brain cortex, kidney)
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    20 µg
    Specification
    Right ventricle, brain cortex, kidney
    Blocking step
    Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

    Annabelle Batten

    Verified customer

    Submitted Oct 23 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (retina)
    Gel Running Conditions
    Non-reduced Denaturing (10 TGX)
    Loading amount
    20 µg
    Specification
    retina
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Feb 12 2018

    Application
    Western blot
    Sample
    Mouse Tissue lysate - other (Isolated mitochondria)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    30 µg
    Specification
    Isolated mitochondria
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C

    Abcam user community

    Verified customer

    Submitted Nov 28 2017

    Application
    Western blot
    Loading amount
    25 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (hepatocytes)
    Specification
    hepatocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

    Abcam user community

    Verified customer

    Submitted Jan 14 2015

    Application
    Western blot
    Loading amount
    20000 cells
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Human Cell lysate - whole cell (jurkats)
    Specification
    jurkats
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

    Dr. Tejpal Gill

    Verified customer

    Submitted Jan 13 2014

    Answer

    Thank you for your reply.

    We have a couple of other catalase antibodies that seem to work well, ab1877 or ab16731-

    https://www.abcam.com/Catalase-antibody-Peroxisome-Marker-ab1877.html

    https://www.abcam.com/Catalase-antibody-ab16731.html

    We have tested this antibodies in liver, placenta, or HeLa lysate and any of them will be appropriate as a positive control. Which would you like to try?

    I look forward to hearing from you. Please let know if you have any questions or need anything else from me.

    Read More

    Answer

    Thank you for contacting us and letting us know about the trouble with this antibody. I am sorry to hear that the results have not been satisfactory.

    I looked through our records and found the previous correspondence you had with my colleague about the first vial of ab52477. Could you confirm that the protocol details are the same as before?

    1) primary antibody dilution and incubation time - 1:1000 dilution and overnight incubation at 4C.
    2) what cell samples used - Used PBT and Jurkat cells
    3) how were the samples treated - Samples were lysed and then boiled
    4) how much protein loaded - 3ug/sample /well
    5) gel electrophoresis conditions - 125 Volts for 90 mins and transferred at 25 volts for 90 mins
    6) transfer and blocking conditions (how long and what reagent was used) - 3% BSA for blocking, Primary and Secondary.
    7) secondary antibody dilution and incubation time - HRP, 1:2000 dilution and 1 hr incubation. Western using other antibodies with the same secondary gave me nice results.
    8) detection method used - super signal west pico chemiluminescent substrate

    I also have a couple of additional questions. What was the largest amount of protein that was loaded per lane of the gel?Was the protein transfer verifiedwith a Ponceau stain?

    We do guarantee all of our products to work as stated on the datasheets for up to 6 months after purchase.I would consider Jurkat cells and PBTs to be positive controls, as I would expect them to beexpressingcatalase, so I would be happy to send you a different antibody if you would like. We also have ab1877, which has been testedwith human cells in Western blot-

    https://www.abcam.com/Catalase-antibody-Peroxisome-Marker-ab1877.html

    I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (Rat cell lines)
    Loading amount
    20 µg
    Specification
    Rat cell lines
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

    Abcam user community

    Verified customer

    Submitted Feb 20 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human cancer cell lines)
    Loading amount
    20 µg
    Specification
    Human cancer cell lines
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

    Abcam user community

    Verified customer

    Submitted Feb 20 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (MEF F9 GC1 NIH-3T3)
    Loading amount
    20 µg
    Specification
    MEF F9 GC1 NIH-3T3
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

    Abcam user community

    Verified customer

    Submitted Aug 02 2010

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

    Sign up