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Superoxide Dismutase Activity Assay Kit (Colorimetric) (ab65354)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1)Q&A (25)References (70)

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Functional studies - ab65354
  • Functional Studies- Superoxide Dismutase Activity Assay Kit (ab65354)
  • Functional Studies - Superoxide Dismutase Activity Assay Kit (Colorimetric) (ab65354)
  • Functional Studies - Superoxide Dismutase Activity Assay Kit (Colorimetric) (ab65354)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 30 min
  • Sample type: Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine

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Overview

  • Product name

    Superoxide Dismutase Activity Assay Kit (Colorimetric)
  • Detection method

    Colorimetric
  • Sample type

    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay type

    Enzyme activity
  • Assay time

    0h 30m
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Superoxide Dismutase Activity Assay Kit (Colorimetric) ab65354 is a simple and rapid assay for superoxide dismutase (SOD) activity.


    In the SOD assay protocol:
    - superoxide anions are produced by the action of xanthine oxidase
    - SOD catalyzes the dismutation of the superoxide anion into hydrogen peroxide and O2
    - superoxide anions act on WST-1 to produce a water-soluble formazan dye which can be detected by the increase in absorbance at 450 nm
    The greater the activity of SOD in the sample, the less formazan dye is produced.


    Superoxide dismutase assay protocol summary:
    - add samples to wells
    - add WST-1 working solution and enzyme working solution and incubate for 20 min at 37ºC
    - analyze with microplate reader

  • Notes

    Superoxide dismutase (SOD) is one of the most important antioxidative enzymes. It catalyzes the dismutation of the superoxide anion into hydrogen peroxide and molecular oxygen.

    Related products

    Review the oxidative stress marker and assay guide to learn about more assays for oxidative stress.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components Identifier 100 tests
    SOD Assay Buffer WM 1 x 20ml
    SOD Dilution Buffer NM 1 x 10ml
    SOD Enzyme Solution Green 1 x 20µl
    WST Solution Red 1 x 1ml
  • Research areas

    • Cancer
    • Cancer Metabolism
    • Cellular metabolic process
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Oxidative stress
    • Metabolism
    • Types of disease
    • Cancer
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Cell Damage
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Antioxidant
  • Relevance

    Superoxide dismutase (SOD) is an antioxidant enzyme involved in the defense system against reactive oxygen species (ROS). SOD catalyzes the dismutation reaction of superoxide radical anion (O2-) to hydrogen peroxide, which is then catalyzed to innocuous O2 and H2O by glutathione peroxidase and catalase. Several classes of SOD have been identified. These include intracellular copper, zinc SOD (Cu, Zn SOD/SOD1), mitochondrial manganese SOD (Mn SOD/SOD2) and extracellular Cu, Zn SOD (EC SOD/SOD3).
  • Cellular localization

    Cytoplasmic
  • Alternative names

    • ALS
    • ALS1
    • Cu /Zn superoxide dismutase
    • Cu/Zn superoxide dismutase
    • IPOA
    • SOD
    • SOD soluble
    • SOD1
    • Superoxide dismutase 1 soluble
    • Superoxide dismutase cystolic
    see all

Associated products

  • Related Products

    • Recombinant human Superoxide Dismutase 1 protein (Active) (ab112193)

Images

  • Functional studies - ab65354
    Functional studies - ab65354Image from Park J et al., PLoS One 12(2), fig 4b. Doi: 10.1371/journal.pone.0172751 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Park J et al investigates the recovery in erectile function after administration of chronic statin alone in DM (streptozotocin (STZ)-induced diabetes mellitus) rats. SOD activity was determined using Superoxide Dismutase activity assay kit (ab65354).

    * Indicates statistical significance in comparison with DM group (P < 0.05). # Indicates statistical significance in comparison with the statin group (P<0.05).

  • Functional Studies- Superoxide Dismutase Activity Assay Kit (ab65354)
    Functional Studies- Superoxide Dismutase Activity Assay Kit (ab65354)

    Superoxide dismutase measured in biofluids at various dilutions

  • Functional Studies - Superoxide Dismutase Activity Assay Kit (Colorimetric) (ab65354)
    Functional Studies - Superoxide Dismutase Activity Assay Kit (Colorimetric) (ab65354)

    Superoxidase dismutase (ab90040) measured showing inhibition rate (%) per concentration (microgram per mL)

  • Functional Studies - Superoxide Dismutase Activity Assay Kit (Colorimetric) (ab65354)
    Functional Studies - Superoxide Dismutase Activity Assay Kit (Colorimetric) (ab65354)
    Principle of Superoxide Dismutase Assay.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (70)

    Publishing research using ab65354? Please let us know so that we can cite the reference in this datasheet.

    ab65354 has been referenced in 70 publications.

    • Shastri S  et al. Idebenone Protects against Acute Murine Colitis via Antioxidant and Anti-Inflammatory Mechanisms. Int J Mol Sci 21:N/A (2020). PubMed: 31940911
    • Li J  et al. CD5L-induced activation of autophagy is associated with hepatoprotection in ischemic reperfusion injury via the CD36/ATG7 axis. Exp Ther Med 19:2588-2596 (2020). PubMed: 32256738
    • Han J  et al. Effects of temperature changes on life parameters, oxidative stress, and antioxidant defense system in the monogonont marine rotifer Brachionus plicatilis. Mar Pollut Bull 155:111062 (2020). PubMed: 32469753
    • Zhou H  et al. Chidamide Inhibits Glioma Cells by Increasing Oxidative Stress via the miRNA-338-5p Regulation of Hedgehog Signaling. Oxid Med Cell Longev 2020:7126976 (2020). PubMed: 32256960
    • Manzoor R  et al. Synergistic Neuroprotective Effect of Endogenously-Produced Hydroxytyrosol and Synaptic Vesicle Proteins on Pheochromocytoma Cell Line Against Salsolinol. Molecules 25:N/A (2020). PubMed: 32276517
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 26 Abreviews or Q&A

    Question

    1. Can we do the test without SOD standard? Do you have some explanation how we should do the test if we dont have the SOD standard

    2. If we should order the SOD how much the price and how long it takes to order.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 20 2014

    Answer

    The ab65354 Superoxide Dismutase Activity Colorimetric Assay Kit can be used without a standard since it is reporting as % inhibition in your sample. Blank 1 does not contain any inhibition, and your sample will have some percentage of inhibition. Blank 2 and Blank 3 are background values so remember to minus these values when doing the calculation. Thus, the more SOD in a sample the less WST-1 formazan produced (which was dependent on Xanthine Oxidase (XO) activity).


    If you want to measure the amount of SOD in your sample, you can use a purified protein as a standard (ab112193 SOD full length protein) and then treat the standard just as you would treat a sample and calculate the amount using a standard curve. Here are the details of the standard.


    For your information, a 96-well plate is also not included in the kit but you can use a standard flat bottom plate such as the one sold by Greiner Bio one (#655-101) or any other standard polystyrene plate.

    Read More

    達偉 鄭

    Abcam Scientific Support

    Answered on Jan 20 2014

    Question

    Is it possible to prolong the shelf life of product ab65354 by storing WST Working Solution in different conditions than the 4C stated in the protocol after reconstitution?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 26 2012

    Answer

    I would not suggest this, as we store this product at 4C ourselves. We haven't tried to store this productat -20C, and we do not know if the product is suitable to be stored at lower temperatures. Please be aware that the customer will loose the Abpromise if they store the product at -20C.

    Read More

    Abcam Scientific Support

    Answered on Mar 26 2012

    Question

    Hi, Can you please tell me what is the lower limit of detection for this assay? Thank you!

    Read More

    Abcam community

    Verified customer

    Asked on May 19 2010

    Answer

    The lower limit for this kit is 0.1 U/ml of SOD activity.

    Read More

    Abcam Scientific Support

    Answered on May 27 2010

    Question

    We calculated the results as inhibition rate%. To calculate the SOD activity in U/ml, should we use the standard or is there any conversion factor for this?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 07 2015

    Answer

    This kit is not originally designed to be run with a standard curve each time. However, if you wish to run a standard curve to calculate absolute activity (the assay is designed to calculate percent inhibition with respect to control sample), you can try using the human recombinant active SOD enzyme

    ab112193
    https://www.abcam.com/Recombinant-human-Superoxide-Dismutase-1-protein-Active-ab112193.html (or use the following: https://www.abcam.com/Recombinant-human-Superoxide-Dismutase-1-protein-Active-ab112193.html).

    Read More

    Sam Washer

    Abcam Scientific Support

    Answered on Sep 07 2015

    Question

    Which one of the three isoforms of SOD is directed for in the kit?

    Read More

    Abcam community

    Verified customer

    Asked on May 20 2014

    Answer

    I can confirm that there is nothing isoform-specific in the kit principle. ab65354 measures the oxygen anion created by any of the SOD enzymes.

    Read More

    Anja Hoffmann

    Abcam Scientific Support

    Answered on May 20 2014

    Question

    What mass of tissue is recommended as starting material for this assay?

    Read More

    Abcam community

    Verified customer

    Asked on Jun 07 2013

    Answer

    The minimum mass of tissue to use with this assay is 10mg.

    Read More

    Abcam Scientific Support

    Answered on Jun 07 2013

    Question

    Could you please advise me, should the protein concentration be measured of the samples tested? The protocol you have provided shows a graph of percent inhibition vs SOD activity, has this been normalised to the protein concentration?

    Read More

    Abcam community

    Verified customer

    Asked on May 15 2013

    Answer

    The graph we have on our datasheet has the % inhibition on Y-axis. From this kit protocol, you will just get the Y-value which is the % inhibition. To draw the graph, you will need the X value. You can get that by using standards (not included in the kit). The standards (ab112193) have to be treated the same way as the samples, but since you already know the amount you are taking, you can easily calculate U/ml.

    Read More

    Abcam Scientific Support

    Answered on May 15 2013

    Question

    How much tissue should be used for extraction?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 15 2013

    Answer

    We would recommend using at least 10 mg of tissue with the kit ab65354.

    Read More

    Abcam Scientific Support

    Answered on Mar 15 2013

    Question

    How do we prepare blood samples to use with this kit?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 11 2013

    Answer

    Collect using citrate or EDTA. Centrifuge at 1,000 x g for 10 min at 4°C. Remove plasma layer w/o disturbing the buffy layer and store at -80°C until ready for analysis. Remove the buffy layer from the red cell pellet. Resuspend erythrocytes in 5x volume of ice cold distilled water and centrifuge at 10,000 x g to pellet the erythrocyte membranes. Store supernatant at -80°C until ready for analysis. Plasma can be diluted approx 3-10x and red cell lysate diluted approx 100x prior to SOD assay.

    Read More

    Abcam Scientific Support

    Answered on Mar 11 2013

    Question

    Are three blanks required for each sample, or can three blanks suffice for comparison to all samples run at the same time?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 11 2013

    Answer

    Yes, you would need the 3 controls with each and every sample.

    Read More

    Abcam Scientific Support

    Answered on Feb 11 2013

    1-10 of 26 Abreviews or Q&A

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