To begin, prepare the test line, and control line coating solution. Dilute the C-reactive protein capture antibody, and Goat anti-rabbit immunoglobulin G with phosphate buffer. Next, place the polyvinyl chloride board properly and remove the protective paper located 25 millimeters from the edge of the board.
Align and adhere the nitrocellulose membrane along the lower edge of the remaining protective paper to the polyvinyl chloride board. Dispense the test line, and control line coating solutions onto the nitrocellulose membrane, ensuring it is placed 20 millimeters apart from the upper edge, forming a test line. For drying, place the nitrocellulose membrane in conditions with a humidity of less than 30%and a temperature range of 18 to 26 degrees Celsius.
To prepare the conjugate pad, submerge the conjugate pad into the conjugate pad treatment solution, then place the soaked conjugate pad in a drying oven for overnight drying treatment. Dilute the prepared QD nano bead antibody conjugate solution with the preservative solution, and dispense it onto the conjugate pad. For sample pad preparation.
Submerge the sample pad into the sample pad treatment solution. Then place the soaked sample pad into a drying oven set at 45 degrees Celsius, and dry for 24 hours. Successively, place the absorbent paper, conjugate pad and sample pad on the polyvinyl chloride board with the nitrocellulose membrane.
Pipette 10 microliters of plasma sample or calibrator solution into one milliliter of A PBS buffer containing 0.1%tween 20. Then transfer 100 microliters of the prepared diluted sample onto the sample pad of the test strip. After a 15 minute run, observe the fluorescent signals of the test line and control line under ultraviolet light exposure to obtain qualitative results.
Insert the strip card into a fluorescent lateral flow assay reader to obtain the fluorescence intensities for quantitative analysis.
