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: Other species, Mammals
  • 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.

  • 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
  • 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

Images

  • 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).

  • Superoxide dismutase measured in biofluids at various dilutions

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

  • Principle of Superoxide Dismutase Assay.

Protocols

References

This product has been referenced in:
  • Cabrejos DAL  et al. Structural characterization of a pathogenicity-related superoxide dismutase codified by a probably essential gene in Xanthomonas citri subsp. citri. PLoS One 14:e0209988 (2019). Read more (PubMed: 30615696) »
  • Kaushal JB  et al. Sonic hedgehog protects endometrial hyperplasial cells against oxidative stress via suppressing mitochondrial fission protein dynamin-like GTPase (Drp1). Free Radic Biol Med 129:582-599 (2018). Read more (PubMed: 30347228) »
See all 35 Publications for this product

Customer reviews and Q&As

1-10 of 26 Abreviews or Q&A

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

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

Question
Answer

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

Read More

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/index.html?datasheet=112193 (or use the following: https://www.abcam.com/index.html?datasheet=112193).

Read More

Question
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

Question
Answer

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

Read More

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

Question
Answer

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

Read More

Question
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

Answer

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

Read More

1-10 of 26 Abreviews or Q&A

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

Sign up