Mitochondrial Complex I (NADH-CoQ Reductase) Activity Assay Kit

Mitochondrial Complex I (NADH-CoQ Reductase) Activity Assay Kit

SKU #	E-BC-K149-M
Detection Instrument	Microplate reader (340 nm)
Detection method	Colorimetric method

Product Details

Properties

Synonyms	Animal tissue
Sensitivity	15.58 U/L
Detection Range	15.58-224.2 U/L
Detection Method	Colorimetric method
Assay type	Enzyme Activity
Assay time	15 min
Precision	Average inter-assay CV: 8.400% | Average intra-assay CV: 0.800%
Other instruments required	Centrifuge, 37℃ incubator
Other reagents required	Anhydrous ethanol (AR)
Storage	-20℃
Valid period	12 months

Detection Principle

Mitochondrial complex I catalyzes the reaction of NADH with ubiquinone substrate to generate NAD+ and reduced ubiquinone. The specific inhibitor rotenone can inhibit the enzyme, and the total enzyme activity minus the enzyme activity after inhibitor inhibition can reflect its specific enzyme activity.

 

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	25 mL × 1 vial	50 mL × 1 vial	2-8°C, 12 months
Reagent 3	Protease Inhibitor	0.4 mL × 1 vial	0.4 mL × 2 vials	2-8°C, 12 months, shading light
Reagent 4	Buffer Solution	15 mL × 1 vial	15 mL × 2 vials	2-8°C, 12 months, shading light
Reagent 5	Substrate A	Powder ×1 vial	Powder ×2 vials	2-8°C, 12 months, shading light
Reagent 6	Substrate B	Powder ×1 vial	Powder ×1 vial	2-8°C, 12 months, shading light
Reagent 7	Inhibitor	1.5 mL × 1 vial	1.5 mL × 2 vials	2-8°C, 12 months, shading light
Reagent 8	Negative Reagent	1.5 mL × 1 vial	1.5 mL × 2 vials	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 lung 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)	32.50	105.00	197.00
%CV	1.0	0.7	0.7

Inter-assay Precision

Three rat lung 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)	32.50	105.00	197.00
%CV	8.2	8.6	8.4

 

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 102%.

	Sample 1	Sample 2	Sample 3
Expected Conc. (U/L)	56.8	135	188
Observed Conc. (U/L)	58.5	132.3	197.4
Recovery rate (%)	103	98	105

 

Sensitivity

The analytical sensitivity of the assay is 15.58 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.