A kind of colorimetric method and kit for detecting ATP
A colorimetric and chromogenic agent technology, applied in the detection field, can solve the problems of expensive instruments, cumbersome operation steps, time-consuming and labor-consuming, etc., and achieve the effect of simple method and wide application prospect
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Embodiment 1
[0032] Accurately pipette 0 μL, 20 μL, 50 μL, 80 μL, 100 μL, 150 μL, and 200 μL of ATP solution with a concentration of 1 mM, add them into the prepared 1.5 mL centrifuge tube, and add 200 μL, 180 μL, 150 μL, 120 μL, 100 μL into the centrifuge tube in turn , 50 μL, 0 μL water, at this time, the volume of the solution in the centrifuge tube is 200 μL. Add 100 μL of 1 mM HO to each centrifuge tube 2 o 2 Solution and 20 μL of ABTS solution with a concentration of 50 mM were added to 680 μL of acetic acid solution with a concentration of 0.1 M, and the volume was adjusted to 1 mL, and reacted at room temperature for 2 hours to obtain the following: figure 1 solutions indicated. The concentration of ATP (from left to right) is 0 μM, 20 μM, 50 μM, 80 μM, 100 μM, 150 μM, 200 μM. It can be seen that with the increase of ATP concentration, the color of the solution becomes lighter and lighter.
[0033] Utilize the ultraviolet-visible spectrophotometer to measure the absorbance of th...
Embodiment 2
[0035] Operation is substantially the same as Example 1, and the difference is only: replace the acetic acid solution of Example 1 with hydrochloric acid solution, and the reaction times are different, and the concrete steps are as follows:
[0036] Accurately pipette 0μL, 20μL, 50μL, 80μL, 100μL, 150μL, 200μL of ATP solution (1mM) into the prepared 1.5mL centrifuge tube, then add 200μL, 180μL, 150μL, 120μL, 100μL, 50μL into the centrifuge tube , 0 μL water, add 100 μL 1mM HO to each centrifuge tube 2 o 2 solution and 20μL 50mM ABTS solution, respectively, with 680μL of 0.1M hydrochloric acid solution to 1mL, react at room temperature for 1 hour to obtain similar figure 1 solutions shown. As the concentration of ATP increases, the color of the solution becomes lighter and the concentration of ATP is 0 μM, 20 μM, 50 μM, 80 μM, 100 μM, 150 μM, and 200 μM, respectively.
[0037] The absorbance of the solution at 380nm to 800nm was measured with a UV-visible spectrophotometer...
Embodiment 3
[0039] Operation is substantially the same as Example 1, and the difference is only: replace the acetic acid solution of Example 1 with phosphoric acid solution, and the reaction times are different, and the concrete steps are as follows:
[0040] Accurately pipette 0μL, 20μL, 50μL, 80μL, 100μL, 150μL, 200μL of ATP solution (1mM) into the prepared 1.5mL centrifuge tube, then add 200μL, 180μL, 150μL, 120μL, 100μL, 50μL into the centrifuge tube , 0 μL water, add 100 μL 1mM HO to each centrifuge tube 2 o 2 Solution and 20μL 50mM ABTS solution, respectively, with 680μL concentration of 0.1M phosphoric acid solution to 1mL, reacted at room temperature for 0.5 hours to obtain similar figure 1solutions shown. As the concentration of ATP increases, the color of the solution becomes lighter and the concentration of ATP is 0 μM, 20 μM, 50 μM, 80 μM, 100 μM, 150 μM, 200 μM.
[0041] The absorbance of the solution at 380nm to 800nm was measured with a UV-visible spectrophotometer, an...
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