Chemical luminescence immune assay determination reagent kit for prostate gland acid phosphatase and preparation method thereof
A technology of phosphatase chemistry and acid phosphatase, applied in the field of clinical blood immunoassay, can solve the problems of narrow detection range, inability to store for a long time, short half-life of radionuclides, etc., and achieves easy popularization and application, no radioactive pollution, and reduced inspection costs. Effect
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Embodiment 1
[0043] Example 1 Preparation of Prostate Acid Phosphatase Chemiluminescence Immunoassay Assay Kit of the present invention
[0044] 1. Preparation of enzyme-labeled prostatic acid phosphatase monoclonal antibody
[0045] 1. Improved glutaraldehyde cross-linking method to label alkaline phosphatase (ALP)
[0046] 1) Take 0.50mg of ALP and 1.0mg of monoclonal antibody, dissolve them in normal saline respectively, mix them, and control the final volume at 0.50ml;
[0047] 2) Add 0.50ml of 1% glutaraldehyde, stir magnetically at room temperature for 15 minutes, and react in the dark for 4 hours;
[0048]3) Add 0.10ml of 1.0mol / L ethanolamine and incubate at room temperature for 2h;
[0049] 4) Under the condition of 4°C, use PBS and magnetic stirring to dialyze overnight, and change the dialysate twice;
[0050] 5) Transfer the dialysis product into a reagent bottle, add an equal volume of glycerin, mix well, add 1% BSA, and store in an airtight freezer for later use.
[0051]...
Embodiment 2~3
[0115] Embodiment 2~3 preparation prostatic acid phosphatase chemiluminescence immunoassay assay kit of the present invention
[0116] The PAP chemiluminescent immunoassay assay kit was prepared in the same manner as in Example 1 except that plastic beads and plastic tubes were used as carriers respectively.
Embodiment 4
[0117] Embodiment 4 The usage method of the kit of the present invention
[0118]Before using this kit for experiments, it is necessary to take out the solid-phase antibody, calibrator / test sample, and labeled antibody solution and place them at room temperature for 15 to 30 minutes to allow them to equilibrate to room temperature; after that, prepare a suitable micro-sampler and corresponding Tips and check whether the chemiluminescence instrument and auxiliary equipment, such as a plate washer, are working properly.
[0119] The specific operation steps of using this kit to carry out the experiment according to the method of embodiment 1 are as follows:
[0120] 1) Place the strips required for the experiment (antibody-coated, 96 / 48 wells can be disassembled into 12 / 6*8*1 wells) on the plate rack;
[0121] 2) Add the sample to be tested and calibrators of each concentration in the reaction wells, add 0, 2, 5, 10, 25, 60 ng / ml of 25 μl of the calibrators to each well, set a ...
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