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Superoxide Dismutase Activity Assay Kit (Colorimetric) (ab65354)

  • Datasheet
  • Protocol Booklet
Reviews (2)Q&A (25)References (116)

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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

    This product is manufactured by BioVision, an Abcam company and was previously called K335 Superoxide Dismutase (SOD) Activity Colorimetric Assay Kit. K335-100 is the same size as the 100 test size of ab65354.

    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 2000 tests
    SOD Assay Buffer WM 1 x 20ml 20 x 20ml
    SOD Dilution Buffer NM 1 x 10ml 20 x 10ml
    SOD Enzyme Solution Green 1 x 20µl 20 x 20µl
    WST Solution Red 1 x 1ml 20 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
    • Neuroscience
    • Diseases
  • 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 download

    Download

References (116)

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

ab65354 has been referenced in 116 publications.

  • Li M  et al. Brachyury engineers cardiac repair competent stem cells. Stem Cells Transl Med 10:385-397 (2021). PubMed: 33098750
  • Zagórska-Dziok M  et al. Cosmetic and Dermatological Properties of Selected Ayurvedic Plant Extracts. Molecules 26:N/A (2021). PubMed: 33503925
  • Zagórska-Dziok M  et al. Positive Effect of Cannabis sativa L. Herb Extracts on Skin Cells and Assessment of Cannabinoid-Based Hydrogels Properties. Molecules 26:N/A (2021). PubMed: 33557174
  • Sun C  et al. Atractylenolide I alleviates ischemia/reperfusion injury by preserving mitochondrial function and inhibiting caspase-3 activity. J Int Med Res 49:300060521993315 (2021). PubMed: 33641489
  • Elmorsy E  et al. The protective effect of Indian Catechu methanolic extract against aluminum chloride-induced neurotoxicity, A rodent model of Alzheimer's disease. Heliyon 7:e06269 (2021). PubMed: 33665436
View all Publications for this product

Customer reviews and Q&As

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Submit a review Submit a question

1-10 of 27 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 27 Abreviews or Q&A

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