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Nitric Oxide (NO) Colorimetric Assay Kit– MSE Supplies LLC

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Nitric Oxide (NO) Colorimetric Assay Kit

SKU: E-BC-K035-S-100

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Nitric Oxide (NO) Colorimetric Assay Kit

SKU # E-BC-K035-S
Detection Instrument Spectrophotometer (550 nm)
Detection Method Colorimetric method


Product Details

Properties

Synonyms NO
Sample Type Serum, plasma, tissue
Sensitivity 0.97 μmol/L
Detection Range 0.97-700 μmol/L
Detection Method Colorimetric method
Assay type Quantitative
Assay time 80 min
Precision Average inter-assay CV: 5.200% | Average intra-assay CV: 3.400%
Other instruments required Vortex mixer, Centrifuge, Analytical Balance, Micropipettor
Other reagents required Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4)
Storage Reagent 6: -20℃, others: 2-8℃
Valid period 12 months


Images

BYWng et al investigate the mechanism between cold exposure, gut microbiota and hypertension. Nitric oxide (NO) level of rat plasma was determined using NO colorimetric assay kit (E-BC-K035-S).

NO level was significantly reduced by cold exposure. (P<0.01)

 

Dilution of Sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.97-700 μmol/L).

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

Sample type Dilution factor
Human serum 1
Human plasma 1
10% Mouse liver tissue homogenization 1
Rat serum 1
Mouse serum 1
10% Epipremnum aureum tissue homogenization 1

 

Note: The diluent is double distilled water, normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).

 

Detection Principle

NO is easily oxidized to form NO2- in vivo or in aqueous solution, and a reddish azo compound is formed with the color developing agent, and the concentration of the azo compound is linearly related to the concentration of NO. The concentration of NO can be calculated indirectly by measuring the OD value at 550 nm.


Kit Components & Storage

Item Component Size 1 (50 assays) Size 2 (100 assays) Storage
Reagent 1 Sulphate Solution 50 mL × 2 vials 50 mL × 4 vials 2-8℃, 12 months
Reagent 2 Alkali Reagent 50 mL × 1 vial 50 mL × 2 vials 2-8℃, 12 months
Reagent 3 Chromogenic
Agent A
19 mL ×1 vial 38 mL ×1 vial 2-8℃, 12 months
shading light
Reagent 4 Chromogenic
Agent B
Powder × 1 vial Powder × 1 vial 2-8℃, 12 months
shading light
Reagent 5 Acid Solution 12.5 mL × 1 vial 25 mL × 1 vial 2-8℃, 12 months
Reagent 6 Sodium Nitrite
Standard
Powder × 1 vial Powder × 2 vials 2-8℃, 12 months

 

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 (μmol/L) 18.50 264.00 457.00
%CV 3.8 3.2 3.2


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 (μmol/L) 18.50 264.00 457.00
%CV 5.0 4.9 5.7


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 99%.


Sample 1 Sample 2 Sample 3
Expected Conc. (μmol/L) 164 395 522
Observed Conc. (μmol/L) 162.4 395.0 511.6
Recovery rate (%) 99 100 98


Sensitivity

The analytical sensitivity of the assay is 0.97 μmol/L NO. 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.