ATPase Activity Assay Kit

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ATPase Activity Assay Kit

SKU #	E-BC-K831-M
Detection Instrument	Microplate reader (630-640 nm, optimum wavelength: 640 nm)
Detection method:	Colorimetric method

Product Details

Properties

Sample type	Animal tissue, cell
Sensitivity	0.06 U/L
Detection range	0.06 - 2.75 U/L
Detection method	Colorimetric method
Assay type	Enzyme Activity
Assay time	80 min
Precision	Average inter-assay CV: 5% \| Average intra-assay CV: 4%
Other instruments required	Vortex mixer, Incubator, Centrifuge
Other reagents required	Double distilled water, Normal saline (0.9% NaCl)
Storage	2-8℃
Valid period	12 months

Dilution of Sample

The recommended dilution factor for different samples is as follows (for reference only):

Sample type	Dilution factor
10% Rat liver tissue homogenate	5-10
10% Rat kidney tissue homogenate	5-10
10% Rat heart tissue homogenate	5-10
10% Rat spleen tissue homogenate	5-10
10% Rat lung tissue homogenate	5-10
10% Mouse liver tissue homogenate	5-10
10% Mouse spleen tissue homogenate	5-10
10% Mouse lung tissue homogenate	5-10
10% Mouse kidney tissue homogenate	5-10
293T cell	1

Note: The diluent is normal saline (0.9%NaCl). For the dilution of other sample types, please do pretest to confirm the dilution factor.

Detection Principle

ATPase catalyzes the hydrolysis of adenosine triphosphate (ATP) to adenosine diphosphate (ADP) and inorganic phosphorus. The activity of ATPase was determined by inorganic phosphorus production per unit time.

Kit Components & Storage

Item	Component	Size (96 T)	Storage
Reagent 1	Buffer Solution	30 mL × 1 vial	2-8℃, 12 months
Reagent 2	Substrate	Powder × 1 vial	2-8℃, 12 months, shading light
Reagent 3	Acid Reagent	25 mL × 1 vial	2-8℃, 12 months
Reagent 4	Chromogenic Agent A	12 mL × 1 vial	2-8℃, 12 months, shading light
Reagent 5	Chromogenic Agent B	4 mL × 1 vial	2-8℃, 12 months
Reagent 6	10 mmol/L Standard	0.5 mL × 1 vial	2-8℃, 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)	0.85	1.36	2.25
%CV	4.4	3.8	3.8

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)	0.85	1.36	2.25
%CV	4.6	5.2	5.2

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 95%.

	Standard 1	Standard 2	Standard 3
Expected Conc. (mmol/L)	0.015	0.04	0.075
Observed Conc. (mmol/L)	0.0	0.0	0.1
Recovery rate(%)	96	98	91

Sensitivity

The analytical sensitivity of the assay is 0.06 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	0.01	0.02	0.03	0.05	0.06	0.08	0.10
Average OD	0.048	0.092	0.148	0.209	0.322	0.382	0.494	0.580
Absoluted OD	0	0.045	0.100	0.161	0.275	0.334	0.447	0.540