Detection kit for prostate specific antigen, detection method and application thereof
A prostate-specific, detection kit technology, which can be used in biological tests, measuring devices, material inspection products, etc., and can solve problems such as cumbersome steps and limitations
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[0050] Equipment: high-speed refrigerated centrifuge (16R, Zhuhai Heima Medical Instrument Co., Ltd.), ultrapure water machine (SybergyUV, Merck Millipore), ultraviolet-visible spectrophotometer (Cary60, Agilent Technologies, USA), fluorescence spectrophotometer ( F-4600, Japan Hitachi High-Tech Corporation), agarose horizontal electrophoresis tank (DYCP-31DN, Beijing Liuyi).
[0051] Reagents used: PSA antibody, SMCC (succinimide-4-(N-maleimide)cyclohexane-1-1 hydroxyester), TCEP (tris(2-carboxyethyl)phosphine), mercapto Modified DNA1, thiol-modified DNA2, MB (DNA is biosynthesized by Bao, MB contains an rA base that can be recognized by ionase and is modified with FAM and BHQ groups).
[0052] 1. Antibody modified DNA
[0053] Antibody and SMCC (suitable molar ratio 1:10-20) were reacted in PBS buffer at room temperature for 2 hours, then purified with filter membrane (molecular weight cut-off 10,000MW), washed three times with PBS buffer; Add 2 μL TCEP (100 mM) to sulfhyd...
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