Zilpaterol immune colloidal gold detection card and preparation method thereof
A technology of colloidal gold and detection card, which is applied in the direction of measuring device, biological test, material inspection product, etc., to achieve the effect of simple method and accurate result.
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Embodiment 1
[0022] figure 1 It is the structural diagram of the immune colloidal gold detection card of zilpaterol 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 zilpaterol of the present invention: among the figure 9 is a PVC glue board; 7 is a sample pad; 8 is a colloidal gold bonding pad, and the colloidal gold bonding pad is covered with Colloidal gold marker of monoclonal antibody; 10 is the coated membrane, that is, nitrocellulose membrane, which is coated with zilpaterol-BSA and goat anti-mouse IgG; 11 is the water-absorbent pad, made of water-absorbent material Made of filter paper.
[0023] 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.
[0024] A nitrocellulose membrane 10 is adhered in the middle of the PVC rubber ...
Embodiment 2
[0027] The preparation of zilpaterol colloidal gold detection card is realized by the following steps:
[0028] (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;
[0029] (2) Antibody pretreatment: centrifuge the zilpaterol 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 Dilute 0.01 mol / L PBS to 1 mg / ml, pass through a 0.22 μm filter membrane;
[0030] (3) Preparation of colloidal gold markers: ta...
Embodiment 3
[0036] Zipaterol minimum detection limit test
[0037] Accurately weigh 0.500 g zilpaterol standard substance, dissolve it in ultrapure water and make up 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 label them 1~4 respectively, add 9 mL, 4 mL, 1 mL, and 0 mL of ultrapure water to the test tubes respectively, and prepare the concentrations of zilpaterol standard solutions as follows: 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 samples and drop them on the zilpaterol test card, and observe the results 5 minutes after adding ...
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