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Mitochondrial Complex II Activity Assay Kit– MSE Supplies LLC

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Mitochondrial Complex II Activity Assay Kit - MSE Supplies LLC

Mitochondrial Complex II Activity Assay Kit

SKU: E-BC-K150-M-96

  • £33100
  • Save £5800

Mitochondrial Complex II Activity Assay Kit

SKU # E-BC-K150-M
Detection Instrument Microplate reader (590-610 nm, optimum wavelength: 600 nm)
Detection method Colorimetric method


Product Details

Properties

Sample type Animal tissue, cell
Sensitivity 0.54 U/L
Detection Range 0.54-17.66 U/L
Detection Method Colorimetric method
Assay type Enzyme Activity
Assay time 15 min
Precision Average inter-assay CV: 5% | Average intra-assay CV: 4%
Other instruments required Centrifuge
Other reagents required Normal saline (0.9% NaCl), PBS(0.01 M，pH 7.4)
Storage 2-8℃
Valid period 12 months


Detection Principle

Coenzyme Q, the catalytic product of mitochondrial complex II, can further reduce 2, 6-dichloroindoxol, which has a characteristic absorption peak at 600 nm. Therefore, the activity of mitochondrial complex II can be quantified by measure the change OD value at 600 nm.

 

Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Extraction Solution A 50 mL × 1 vial 50 mL × 2 vials 2-8°C, 12 months
Reagent 2 Extraction Solution B 15 mL × 1 vial 30 mL × 1 vial 2-8°C, 12 months
Reagent 3 Inhibitor 0.8 mL × 1 vial 0.8 mL × 2 vials 2-8°C, 12 months, shading light
Reagent 4 Buffer Solution 10 mL×1 vial 20 mL×1 vial 2-8°C, 12 months
Reagent 5 Substrate A 0.8 mL×1 vial 1.6 mL×1 vial 2-8°C, 12 months, shading light
Reagent 6 Substrate B 1.5 mL×1 vial 1.5 mL×2 vials 2-8°C, 12 months, shading light
Reagent 7 Substrate C 0.6 mL×1 vial 1.2 mL×1 vial 2-8°C, 12 months, shading light
UV 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 rat heart tissue 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 (U/L) 2.35 12.60 15.50
%CV 4.3 3.8 3.9


Inter-assay Precision

Three rat heart tissue samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters Sample 1 Sample 2 Sample 3
Mean (U/L) 2.35 12.60 15.50
%CV 4.9 5.2 4.9

 

Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 97%.

Sample 1 Sample 2 Sample 3
Expected Conc. (μmol/L) 4.5 9.8 14.5
Observed Conc. (μmol/L) 4.5 9.6 13.3
Recovery rate (%) 101 98 92

 

Sensitivity

The analytical sensitivity of the assay is 0.54 U/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.