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Inhibition and Production of Superoxide Anionic Colorimetric Assay Kit– MSE Supplies LLC

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Inhibition and Production of Superoxide Anionic Colorimetric Assay Kit (WST-1 Method) - MSE Supplies LLC

Inhibition and Production of Superoxide Anionic Colorimetric Assay Kit (WST-1 Method)

SKU: E-BC-K001-M-500

  • $ 71495
  • Save $ 12600

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%