Immune colloidal gold detection card for phenylethanolamine A and preparation method thereof
A technology of phenylethanolamine and colloidal gold, applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effect of accurate results and simple methods
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Embodiment 1
[0021] figure 1 It is the structural diagram of the immune colloidal gold detection card of phenylethanolamine A of the present invention, figure 2 It is the cross-sectional structure diagram of the test paper strip in the immune colloidal gold detection card of phenylethanolamine A of the present invention: among the figure 9 is a PVC rubber sheet; 7 is a sample pad; 8 is a colloidal gold bonding pad, and the colloidal gold bonding pad is coated Monoclonal antibody colloidal gold marker; 10 is the coated membrane, that is, nitrocellulose membrane, which is coated with phenylethanolamine A-BSA and goat anti-mouse IgG; 11 is the absorbent pad, made of absorbent material such as filter paper production.
[0022] On one end (sample end) of the PVC rubber plate 9, the sample pad 7 and the colloidal gold bonding pad 8 are adhered, and the sample pad 7 and the colloidal gold bonding pad 8 are in a side-by-side structure.
[0023] A nitrocellulose membrane 10 is adhered in the mid...
Embodiment 2
[0026] The preparation of the phenylethanolamine A colloidal gold detection card is specifically realized by the following steps:
[0027] (1) Preparation of colloidal gold solution: Take a 1 L Erlenmeyer flask, add 495 mL of ultrapure water, and then add 1% chloroauric acid (HAuCl 4 ·3H 2 (0) 5 mL, prepared into 500 mL 0.01% chloroauric acid aqueous solution, heated and boiled, then added 1% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) Solution 5-7 mL, continue to stir and heat, when the color of the solution turns completely transparent purple red, stop heating after maintaining for 5 min, add water to the original volume, cool to room temperature, and store at 2-8°C for later use;
[0028] (2) Antibody pretreatment: Centrifuge the phenylethanolamine A antibody to be labeled at 1000 r / min, 4°C for 20 min, take the supernatant, and dilute it to 1 mg / mL with 0.01 mol / L PBS; or use 0.01 mol / L PBS was diluted to 1 mg / mL, and passed through a 0.22 μm filter membrane;
[002...
Embodiment 3
[0035] Minimum detection limit test
[0036] Phenylethanolamine A minimum detection limit test
[0037] Accurately weigh 0.500 g of phenylethanolamine A standard, dissolve it in ultrapure water and dilute to 100 mL, marked as A 1 solution, take A 1 Add 1 mL of the solution to a volumetric flask and add ultrapure water to 1000 mL to form A 2 solution, take A 2 Add 1 mL of the solution to a volumetric flask and add ultrapure water to 1000 mL to form A 3 solution, the concentration of A3 solution is 5 ng / mL (5ppb). take A 3 Put 1 mL of the solution in 4 test tubes, and mark them as 1~4 respectively, add 9 mL, 4 mL, 1 mL, and 0 mL of ultrapure water to the test tubes respectively, and make the concentration of phenylethanolamine A standard solution: 0.5 ng / mL, 1 ng / mL, 2.5 ng / mL, 5 ng / mL; take another empty test tube, add 5 mL of ultrapure water, and mark it as No. 5 test tube. Take the liquid in the above test tubes 1-5 as a sample and add it dropwise to the phenylethanola...
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