Application of protocatechuic acid as inhibitor of calcium ion channel
A technology of calcium ion channel and protocatechuic acid, which is applied in the application field of protocatechuic acid, can solve the problems that there is no research showing the inhibitory effect of protocatechuic acid
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[0073] (1) Transfect the cDNA plasmid vector containing TRPC3, TRPC6 or TRPC7 ion channels into the HERK293 cell line, and then select the corresponding antibiotics to incubate the cells according to the resistance of the plasmid to selectively screen the successfully transfected cells. Functional tests were performed on the surviving cells to confirm the expression and function of the channel protein, and then cloned and purified through the "limiting dilution" process to obtain a stable cell line that stably expresses a specific channel.
[0074] (2) Add 10-13×104 cells / mL of the stable cells obtained in step (1), and add 150 μL of cell suspension per well into a 96-well plate with a black wall bottom. After 24 hours of cultivation in the incubator, it can be used for subsequent experiments.
[0075] (3) Observe the cells, and after confirming that the cells are in good condition, load the dye (fluo-4) for 60 min.
[0076] (4) prepare channel activator and inhibitor solutio...
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