Storage 

Stored at -20℃ ,The kit is stable for at least one year from the date it is received. NADPH, Nitrate Reductase, NaNO2, Griess reagent I and Griess reagent II should be stored in dark. The solution of NADPH should be divided into several parts and stored at -70℃ . 

Description 

As an important gas signal molecule, NO widely exists in organisms and various tissues, and participates in many biological effects and physiological and pathological processes. NO has free radicals, which makes its chemical properties very active, and rapidly oxidizes to form the mixture of nitrite and nitrate. Based on the principle of Griess reaction, this kit adopts the optimized reaction conditions to reduce nitrate to nitrite by nitrate reductase, so as to achieve the purpose of detecting the content of total NO. 

The kit can detect the content of NO in cell lysate, tissue lysate, cell or tissue culture medium, serum, plasma or urine. 

Notes 

1. If Griess reagent I contains precipitate when taken out, it can be placed in a 37 ℃ water bath until the precipitate dissolves. 

2. For the culture medium containing high concentration of nitrate, it is necessary to replace it with other appropriate medium, such as DMEM, MEM, F12, etc., or HBSS or PBS, etc. before detecting NO. Because higher concentration of nitrate will interfere with the result. 

3. Due to the reduction reaction in the detection process, all oxidation or reduction reagents that affect the reduction reaction should be avoided, such as common reducing agents DTT and mercaptoethanol. 

4. The reaction must be conducted away from light, and bubbles should be avoided in each detection hole when adding reagent each time, so as not to interfere with the test results. 

5. The upper limit of sample dosage is 60 μL, and 40 μL is usually enough for serum, plasma or tissue homogenate. When the sample is less than 60 μL, the volume of different samples should be consistent, and the insufficient volume should be supplemented with the solution used for preparing or diluting the sample. 

6. 2-3 wells with 200 μL water or PBS can be set as negative control at the same time. Only water or PBS is added in these 2-3 holes, and no other reagents are added. 

7. When testing, if there is no 540 nm filter, 520-560 nm filter can also be used.