Inhibition and Production of Superoxide Anionic Colorimetric Assay Kit (WST-1 Method)
SKU: E-BC-K001-M-500
Inhibition and Production of Superoxide Anionic Colorimetric Assay Kit (WST-1 Method)
SKU # | E-BC-K001-M |
Detection Instrument | Microplate reader(440-460 nm, optimum wavelength: 450 nm) |
Detection Method | Colorimetric method |
Product Details
Properties
Sample Type | Serum, plasma, urine, cells, cell culture supernatant, leucocyte |
Assay Time | 35 min |
Precision | Inter-assay CV: 5.8% | intra-assay CV: 2% |
Other Reagents Required | Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
Storage | This product can be stored at 2~8°C/-20°C for 12 months with shading light. Please refer to the manual for the specific storage condition of the components. |
Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
Sample type | Dilution factor |
---|---|
Human serum | 4-7 |
Mouse serum | 15-25 |
Rat serum | 25-35 |
Human saliva | 1 |
HepG2 culture supernatant | 1 |
10% Rat brain tissue homogenate | 150-200 |
10% Rat liver tissue homogenate | 500-600 |
10% Mouse liver tissue homogenate | 500-600 |
10% Mouse heart tissue homogenate | 150-200 |
10% Epipremnum aureum tissue homogenate | 20-30 |
The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Superoxide anion free radicals are produced through the reaction system of xanthine and xanthine oxidase. WST-1 (a water-soluble tetrazolium salt) can react with the generated superoxide anion to produce water-soluble formazan. When the tested sample contains the superoxide anion free radical inhibitor, it can inhibit the formation of formazan. When the tested sample contains the substance that produces superoxide anion free radical, it can promote the formation of formazan dye. By colorimetric analysis of WST-1 products, the units of activity of inhibition or production of superoxide anion radical in samples can be calculated.
Kit Components & Storage
Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
Reagent 1 | Buffer Solution | 12 mL ×1 vial | 24 mL ×1 vial | 2-8℃, 12 months |
Reagent 2 | Substrate Solution | 0.07 mL ×1 vial | 0.14 mL × 1 vial | 2-8℃, 12 months shading light |
Reagent 3 | Enzyme Stock Solution | 0.15 mL ×1 vial | 0.3 mL × 1 vial | -20℃, 12 months |
Reagent 4 | Enzyme Diluent | 1.5 mL ×1 vial | 1.5 mL × 2 vials | -20℃, 12 months |
Reagent 5 | VC Standard | Powder ×3 vials | Powder ×3 vials | 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:
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 (mg/mL) | 0 | 0.0125 | 0.015 | 0.035 | 0.075 | 0.125 | 0.3 | 0.5 |
OD of control | 0.567 | 0.411 | 0.386 | 0.311 | 0.218 | 0.167 | 0.202 | 0.31 |
0.541 | 0.426 | 0.376 | 0.324 | 0.214 | 0.172 | 0.197 | 0.308 | |
OD of sample | 0.039 | 0.039 | 0.041 | 0.042 | 0.052 | 0.064 | 0.15 | 0.252 |
0.039 | 0.04 | 0.041 | 0.043 | 0.049 | 0.065 | 0.145 | 0.251 | |
Average OD of control | 0.554 | 0.419 | 0.381 | 0.318 | 0.216 | 0.17 | 0.2 | 0.309 |
Average OD of sample | 0.039 | 0.04 | 0.041 | 0.043 | 0.051 | 0.065 | 0.148 | 0.252 |
Absolute OD | 0.515 | 0.379 | 0.34 | 0.275 | 0.166 | 0.105 | 0.052 | 0.058 |
Inhibition ratio | 0 | 26% | 34% | 47% | 68% | 80% | 90% | 89% |