Chemiluminescence immunodetection kit and detection method for ovarian cancer tumor marker HE4
A chemiluminescent immune and tumor marker technology, applied in the field of biological and medical detection, can solve the problems of non-detection and achieve good linear range, high affinity, curative effect observation and reliable prognosis judgment
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Embodiment 1
[0030] The ovarian cancer tumor marker HE4 chemiluminescent immunoassay kit provided in this example includes a 9F3 antibody-coated microwell plate, an alkaline phosphatase-labeled 10E1 antibody, an analysis buffer, a substrate working solution, a luminescent substrate, Washing solution, quality control products.
[0031] The preparation method of the 9F3 antibody-coated microwell plate is as follows: dilute the 9F3 antibody to 0.1 μg / μl with coating buffer to prepare the 9F3 antibody coating solution, and then add 50 μg / μl to each well of the microwell plate. Microliter of 9F3 antibody coating solution was coated at 37°C for 2 hours, then the microplate was rinsed three times with normal saline, and then 100 microliter of sealing solution was added to each well of the microplate, and kept at room temperature After blocking for 2 hours, the microwell plate was rinsed twice with normal saline, and freeze-dried to prepare a 9F3 antibody-coated microwell plate, which was sealed i...
Embodiment 2
[0053] The chemiluminescent immunoassay method for the ovarian cancer tumor marker HE4 provided in this example adopts the kit provided in Example 1 of the present invention, and includes the following steps:
[0054] (1) Add BSA to the PBS buffer to prepare a PBS buffer with a BSA mass percentage concentration of 1% BSA, dilute the purchased recombinant HE4 protein with this buffer, and prepare a recombinant HE4 protein standard with a series of concentration gradients solution;
[0055] (2) Take 50 microliters of the human serum to be tested and 50 microliters of the recombinant HE4 protein standard solution with a series of concentration gradients, add them to different wells of the 9F3 antibody-coated microwell plate, and then add the ALP-labeled 10E1 antibody Working solution (1:2000) 50 microliters, then add luminescent substrate CSPD, shake evenly, incubate at 37°C for 1 hour, wash 5 times with washing solution, dry on absorbent paper, then add substrate working solutio...
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