Human epididymis protein immunoassay kit and using method thereof
A human epididymis protein and immune detection technology, applied in the field of biological materials and detection, can solve the problem of low sensitivity, achieve low detection limit, low cost, and improve the effectiveness of detection
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[0026] 1. Preparation of the substrate
[0027] 1. Preparation of immune gold nanoparticles: first prepare gold nanoparticles with a particle size of 35nm, take 1 mL of 0.01 g / mL HAuCl 4(1%) prepared into 100 mL 1.0×10 -4 g / mL HAuCl 4 (0.01%), heated and boiled under vigorous stirring, quickly added 1mL 1.0×10 -2 g / mL sodium citrate aqueous solution, keep boiling for 15 min, then cool to room temperature while stirring. Add 2 μL of 4-MBA labeling molecules, and then add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) and N-hydroxysuccinimide ( NHS) (volume ratio of EDC and NHS: 2:1) solution activation, then inject HE4 antibody, incubate for 2-3 h, centrifuge again, disperse and add 10 μL of 5% (mass volume ratio) BSA to block the remaining active sites.
[0028] 2. Fe 3 o 4 Synthesis of nanoparticles: 0.65 g FeCl 3 ·6H 2 O and 0.2 g of sodium citrate were added to 20 mL of ethylene glycol (EG), and stirred evenly by ultrasonic. 1.2 g of sodium...
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