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Stock availability
In Stock
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Kit s description
Quantitative colorimetric measurement of superoxide dismutase activity
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Protein target
SOD
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Scientific context
Short-lived and highly reactive oxygen species (ROS) such as O2·- (superoxide), ·OH (hydroxyl radical), and H2O2 (hydrogen peroxide) are continuously generated in vivo. In the resting state, the balance between antioxidants and oxidants is sufficient to prevent the disruption of normal physiologic functions; however, either increases in oxidants or decreases in antioxidants can disrupt this balance giving rise to elevated levels of reactive oxygen species (ROS) (1,2).The cellular levels of ROS are controlled by antioxidant enzymes and small molecule antioxidants. The major antioxidant enzymes, superoxide dismutases (SODs), including copper-zinc superoxide dismutase (Cu/ZnSOD, SOD1), manganese superoxide dismutase (MnSOD, SOD2) and extracellular superoxide dismutase (EC-SOD, SOD3), all play critical roles in scavenging O2·-. Decreased SOD activity results in elevated level of superoxide which in turn leads to decreased NO but increased peroxynitrite concentrations. The major intracellular SOD is a 32-kD copper and zinc containing homodimer (Cu/Zn SOD). The mitochondrial SOD (MnSOD) is a manganese-containing 93-kD homotetramer that is synthesized in the cytoplasm and translocated to the inner matrix of mitochondria. EC-SOD is the primary extracellular SOD enzyme and is highly expressed in many organs. Increased SOD activity levels are seen in Downs Syndrome (3) while decreased activity is seen in diabetes, Alzheimer’s disease, rheumatoid arthritis, Parkinson’s disease, uremic anemia, atherosclerosis, some cancers, and thyroid dysfunction (3-8).
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Primary research area
Cancer, Oxidative Stress, Cardiovascular System, Atherosclerosis, Neuroscience, Neurodegeneration, Alzheimer's Disease, Parkinson's Disease
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Category
Assay Activity Kits
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Other name
Cu/ZnSOD Activity Kit, SOD1 Activity Kit, MnSOD Activity Kit, SOD2 Activity Kit, EC-SOD Activity Kit, SOD3 Activity Kit
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Brand name
StressXpress®
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Detection system
Colorimetric Assay
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Assay format
Direct Enzyme Activity Assay
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Main intended usage
Colorimetric assay used to quantitatively measure SOD activity in a variety of samples.
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Brief protocol
The Superoxide Dismutase (SOD) Activity Kit is designed to quantitatively measure SOD activity in a variety of samples. The assay measures all types of SOD activity, including Cu/Zn, Mn, and FeSOD types. A bovine erythrocyte SOD standard is provided to generate a standard curve for the assay and all samples should be read off of the standard curve. Samples are diluted in our specially colored Sample Diluent and added to the wells. The Substrate is added followed by Xanthine Oxidase Reagent and incubated at room temperature for 20 minutes. The Xanthine Oxidase generates superoxide in the presence of oxygen, which converts a colorless substrate in the Detection Reagent into a yellow colored product. The colored product is read at 450 nm. Increasing levels of SOD in the samples causes a decrease in superoxide concentration and a reduction in yellow product. The activity of the SOD in the sample is calculated after making a suitable correction for any dilution, using software available with most plate readers. The results are expressed in terms of units of SOD activity per mL.
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Kit contents
Clear 96 well Half Area Plates, Superoxide Dismutase Standard, Assay Buffer, Xanthine Oxidase Buffer, Xanthine Oxidase Concentrate, Substrate Diluent, Substrate Concentrate
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Assay precision
Intra Assay Precision: Three samples diluted in Assay Buffer were run in replicates of 20 in an assay. The mean and precision of the calculated concentrations were: Sample 1- 0.407 U/mL, 4.6% CV Sample 2- 0.726 U/mL, 7.3% CV Sample 3- 1.203 U/mL, 16.8% CV Inter Assay Precision: Three samples diluted in Assay Buffer were run in duplicates in sixteen assays run over multiple days by four operators. The mean and precision of the calculated concentrations were: Sample 1- 0.356 U/mL, 10.5% CV Sample 2- 0.653 U/mL, 6.1% CV Sample 3- 1.277 U/mL, 13.8% CV
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Assay cross reactivity
Species Independent
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Samples to be used with this kit
Serum, Heparin Plasma, EDTA Plasma, Cell Lysates, Tissue, Erythrocytes
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Maximum samples to be used with this kit
89 samples in duplicate
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Assay duration
See product datasheet or feel free to contact us
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Kit s sensitivity
0,044 U/ml
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Assay detection limit
0,0625 - 4 U/ml
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Storage recommendations
4°C
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Shipping recommendations
Blue Ice
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Bibliography
1. Liocher, SI and Fridovich, I. Free Rad. Biol. Med., 2007, 42:1465-1469. 2. Imlay, JA. Ann Rev. Bichem., 2008, 77:755-776. 3. Torsdottir, G. et al. J.Neurol.Sci. 2010. 299(1-2):51-54. 4. Giacco, F & Brownlee, M. Circ.Res. 2010. 107(9):1058-1070. 5. Bae, S-C, et al. J.Amer.Coll.Nutr. 2003. 22(4):311-315. 6. Akbostanci, MC, et al. Acta Neurol.Belg. 2001. 101:180-183. 7. Shainkin-Kestenbaum, R, et al. Nephron. 1990. 55(3):251-253. 8. Saito, T. Hokkaido Igaku Zasshi. 1987. 62(2):257-268.
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Release date
17-Sep-2014
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PubMed number
Refer to PubMed
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Tested applications
No data available / currently testing
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Tested reactivity
No data available / currently testing
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Resources available upon request
Kit Booklet, MSDS
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Product image link
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Product image legend
Typical Standard Curve for SOD Activity Kit (Enzyme Activity Assay) StressXpress® - SKT-214. Assay Type: Direct Enzyme. Detection Method: Colorimetric Assay. Assay Range: 0.0625 - 4 U/ml. | Linearity was determined by taking two samples, one with a high known SOD activity and the other with a lower SOD activity and mixing them in given ratios. The measured activities were compared to the expected values based on the ratios used. | Short-lived and highly reactive oxygen species (ROS) such as O2·-(superoxide), ·OH (hydroxyl radical), and H2O2 (hydrogen peroxide) are continuously generated in vivo. The cellular levels of ROS are controlled by antioxidant enzymes and small molecule antioxidants. The major antioxidant enzymes, superoxide dismutases (SODs), including copper-zinc superoxide dismutase (Cu/ZnSOD, SOD1), manganese superoxide dismutase (MnSOD, SOD2) and extracellular superoxide dismutase (EC-SOD, SOD3), all play critical roles in scavenging O2·-. Graph of the Typical Standard Curve for the SOD Activity Kit StressXpress - SKT-214 | Graph of the Linearity Recovery for the SOD Activity Kit StressXpress - SKT-214 | Diagram of the Reactive Oxygen Species (ROS) Formation for the SOD Activity Kit StressXpress - SKT-214
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Warnings
Non-hazardous
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Country of production
Canada
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Total weight kg
2
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Net weight g
300
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