Laminin chemiluminescence immune analysis determination reagent kit and preparing method thereof
A chemiluminescence immunoassay and laminin technology, applied in the field of immunoassay medicine, can solve the problems of inability to widely apply clinical diagnosis and scientific research work, needs, and limited popularization and use, to ensure sensitivity, increase sensitivity, and facilitate operation and production. Effect
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Embodiment 1
[0037] Example 1 Preparation of the laminin chemiluminescence immunoassay assay kit of the present invention
[0038] 1. Preparation of enzyme-labeled antibody and preparation of biotin-conjugated antibody
[0039] 1. The polyclonal antibody or monoclonal antibody of laminin is coupled with horseradish peroxidase by periodic acid oxidation method, fully dialyzed against PBS, added an equal volume of glycerol, and stored below -20°C for use.
[0040] 2. Preparation of biotin-conjugated laminin monoclonal antibody
[0041] (1) Use conventional methods to dissolve biotin (BNHS) in N,N-dimethylformamide (DMF) to make 1mg / mL;
[0042] (2) Use 0.1mol / L NaHCO with a pH of 9.0 3 Dilute the purified laminin monoclonal antibody to 1~2mg / mL;
[0043] (3) Mix according to the volume ratio of BNHS and antibody of 1:8 or weight ratio of 1:7, and react for 2 to 4 hours under stirring at room temperature;
[0044] (4) Load the dialysis to 0.05mol / L PBS with a pH of 7.2, dialyze overnight at 4°C, ...
Embodiment 2~3
[0096] Examples 2 to 3 Preparation of the laminin chemiluminescence immunoassay assay kit of the present invention
[0097] Except that plastic beads and plastic tubes are used as carriers, the rest are prepared in the same manner as in Example 1 to prepare a laminin chemiluminescence immunoassay assay kit.
Embodiment 4
[0098] Example 4 Preparation of the laminin chemiluminescence immunoassay assay kit of the present invention
[0099] In addition to using the glutaraldehyde method to label the laminin monoclonal antibody with alkaline phosphatase, using AMPPD as the chemiluminescent substrate, using plastic beads as the carrier, the coating solution is a citrate buffer with a pH of 4.5, and a blocking solution. Except for the pH value of 7.5, the rest were prepared using the same method as in Example 1 to prepare a laminin chemiluminescence immunoassay assay kit.
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