Total Oxidant Status (TOS) Colorimetric Assay Kit
Total Oxidant Status (TOS) Colorimetric Assay Kit
| SKU # | E-BC-K802-M |
| Detection Instrument | Microplate reader (580 nm-590 nm, optimum wavelength: 590 nm) |
| Detection method: | Colorimetric method |
Product Details
Properties
| Synonyms | TOS |
| Sample type | Cell, tissue, serum and other liquid samples |
| Sensitivity | 2.5 μmol H2O2 Equiv./L |
| Detection range | 2.5-100 μmol H2O2 Equiv./L |
| Detection method |
Colorimetric method |
| Assay type | Quantitative |
| Assay time | 30 min |
| Precision | Average inter-assay CV: 3.500% | Average intra-assay CV: 2.300% |
| Other instruments required | Micropipettor, 37℃ incubator. |
| Other reagents required | Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Images
X Zhao et al investigate the function of quercetin for reducing ethanol-induced hepatocyte pyroptosis. Total oxidants status (TOS) of human L02 cell was determined using TOS colorimetric assay kit (E-BC-K802-M).
Different subscript letters indicate significant differences among the groups. (P<0.05)
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (2.5-100 μmol H2O2 Equiv./L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Porcine serum | 1 |
| Horse serum | 1 |
| Cynomolgus macaques serum | 1 |
| 10% Rat liver tissue homogenate | 1 |
| 10% Rat kidney tissue homogenate | 1 |
| 10% Rat lung tissue homogenate | 1 |
| 10% Rat spleen tissue homogenate | 1 |
|
0.612×10^6 Molt-4 cells |
1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
Under acid conditions, the oxidizing material in the sample can oxidize Fe2+ to Fe3+, which binds highly with xylenol orange to produce a blue-purple complex. When the pH of solution is in the range of 2-3, its maximum absorption wavelength is around 590 nm, and the color depth is proportional to the content of oxidation substances in a certain concentration and a certain time, so as to indirectly calculate the total oxidation state of the sample.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Chromogenic Agent | 12 mL × 1 vial | 24 mL × 1 vial | 2-8℃, 12 months, shading light |
| Reagent 2 | Substrate | 3 mL × 1 vial | 6 mL × 1 vial | 2-8℃, 12 months, shading light |
| Reagent 3 | 200 μmol/L H2O2 Standard |
1 mL × 1 vial | 1 mL × 1 vial | 2-8℃, 12 months, shading light |
| Microplate | 96 wells | No requirement | ||
| Plate Sealer | 2 pieces | |||
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol H2O2 Equiv./L) | 5.60 | 34.80 | 85.00 |
| %CV | 2.6 | 2.2 | 2.1 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol H2O2 Equiv./L) | 5.60 | 34.80 | 85.00 |
| %CV | 3.2 | 3.6 | 3.7 |
Sensitivity
The analytical sensitivity of the assay is 2.5 μmol H2O2 Equiv./L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.
Standard Curve
As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:
| Concentration (μmol/L) | 0 | 10 | 20 | 40 | 50 | 60 | 80 | 100 |
| Average OD | 0.085 | 0.146 | 0.195 | 0.337 | 0.439 | 0.502 | 0.668 | 0.767 |
| Absoluted OD | 0.000 | 0.061 | 0.110 | 0.252 | 0.354 | 0.417 | 0.584 | 0.682 |