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α-Ketoglutarate Dehydrogenase(α-KGDH) Activity Assay Kit– MSE Supplies LLC

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α-Ketoglutarate Dehydrogenase(α-KGDH) Activity Assay Kit - MSE Supplies LLC

α-Ketoglutarate Dehydrogenase(α-KGDH) Activity Assay Kit

SKU: E-BC-K083-M-96

  • £24800
  • Save £4400

α-Ketoglutarate Dehydrogenase(α-KGDH) Activity Assay Kit

SKU # E-BC-K083-M
Detection Instrument Microplate reader (440-460 nm, optimum wavelength: 450 nm)
Detection Method Colorimetric method


Product Details

Properties

Synonyms α-KGDH
Sample Type Serum, plasma, animal and plant tissue
Sensitivity 8.3 U/L
Detection Range 8.3-42.3 U/L
Detection Method Colorimetric method
Assay type Enzyme Activity
Assay time 40 min
Precision Average inter-assay CV: 8.500% | Average intra-assay CV: 1.500%
Other instruments required Incubator, Centrifuge
Storage 2-8℃
Valid period 12 months

 

Detection Principle

α-ketoglutarate dehydrogenase (α-KGDH) is a key enzyme in the tricarboxylic acid (TCA) cycle. While the substrate being catalyzed by α-KGDH, NAD+ is reduced to NADH, and electrons are transferred to WST-8 under the action of hydrogen transmitter to produce yellow product. The activity of α-KGDH can be calculated by measuring the change of absorbance value at 450 nm.

 

Kit Components & Storage

Item Component Size 1(96 T) Storage
Reagent 1 Extraction Solution 55 mL ×1 vial -20℃, 12 months
Reagent 2 Buffer Solution 28 mL × 1 vial -20℃, 12 months
Reagent 3 Substrate Powder × 1 vial -20 ℃, 12 months, shading light
Reagent 4 Chromogenic Agent 3 mL × 1 vial -20 ℃, 12 months, shading light
Reagent 5 Clarificant 3 mL × 1 vial -20℃, 12 months
Reagent 6 Standard Powder × 1 vial -20℃, 12 months

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 (U/L) 12.50 27.60 33.80
%CV 1.7 1.4 1.4


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 (U/L) 12.50 27.60 33.80
%CV 8.2 8.5 8.8

 

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 100%.


 Sample 1 Sample 2 Sample 3
Expected Conc. (mmol/L) 0.15 0.28 0.37
Observed Conc. (mmol/L) 0.2 0.3 0.4
Recovery rate (%) 101 98 101


Sensitivity

The analytical sensitivity of the assay is 8.3 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.

 

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 (mmol/L) 0 0.1 0.2 0.25 0.3 0.35 0.4 0.5
Average OD 0.091 0.238 0.436 0.545 0.601 0.681 0.811 1.017
Absoluted OD 0 0.147 0.345 0.453 0.509 0.589 0.719 0.925