SOD Colorimetric Activity Kit

Référence OKAU00026

Conditionnement : 2plate

Marque : Aviva Systems Biology

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Datasheets/ManualsPrintable datasheet for SOD Colorimetric Activity Kit (OKAU00026)
Product Info
ApplicationAA
ELISA Kit Detection MethodColorimetric
ELISA Kit LinearityLinearity 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 the ratios given below. The measured activities were compared to the expected values based on the ratios used.
High SampleLow SampleExpected Activity (U/mL)Observed Activity (U/mL)% Recovery
80%20%0.9870.92093.2
60%40%0.7890.77698.4
40%60%0.5900.53490.4
20%80%0.3920.414105.6
Mean Recovery96.9%
ELISA Kit PrincipleThe DetectX® 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. Please read the complete kit insert before performing this assay. 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 Assay Buffer 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.
ELISA Kit ReproducibilityIntra 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:
SampleSOD Activity (U/mL)%CV
10.4074.6
20.7267.3
31.20316.8
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:
SampleSOD Activity (U/mL)%CV
10.35610.5
20.6536.1
31.27713.8
ELISA Kit Component
ComponentQuantity
Clear 96 well Half Area Plates2 Plates
Superoxide Dismutase Standard1 Vial
Assay Buffer50 mL
Xanthine Oxidase Buffer6 mL
Xanthine Oxidase Concentrate225 uL
Substrate Diluent12 mL
Substrate Concentrate1.1 mL
Additional InformationBackground: 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). 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 while decreased activity is seen in diabetes, Alzheimer's disease, rheumatoid arthritis, Parkinson's disease, uremic anemia, atherosclerosis, some cancers, and thyroid dysfunction.
Reconstitution and Storage2°C to 8°C
Sample TypeSerum, Plasma, Cells, Tissues and Erythrocyte Lysates
Sensitivity0.044 U/mL
Gene SymbolSOD