Cyproheptadine hydrochloride immune colloidal gold detection card and preparation method thereof
The technology of cyproheptadine hydrochloride and colloidal gold is applied in the field of immunocolloidal gold detection card of cyproheptadine hydrochloride and its preparation field, and the effects of accurate results and simple method are achieved.
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Embodiment 1
[0020] figure 1 , figure 2 Shown embodiment: among the figure 9 is a PVC rubber sheet; 7 is a sample pad; 8 is a colloidal gold binding pad, and the colloidal gold binding pad is coated with a monoclonal antibody colloidal gold marker; 10 is a coating film, i.e. nitrocellulose Plain membrane, the nitrocellulose membrane coated with cyproheptadine hydrochloride-BSA and goat anti-mouse IgG; 11 is a water-absorbent pad, made of water-absorbent materials such as filter paper.
[0021] On one end (sample end) of the PVC rubber plate 9, the sample pad 7 and the bonding pad 8 are adhered, and the sample pad 7 and the bonding pad 8 are in a side-by-side structure.
[0022] A nitrocellulose membrane 10 is adhered in the middle of the PVC rubber sheet 9 . Goat anti-mouse IgG quality control line 3 and cyproheptadine hydrochloride-BSA detection line 2 are set on the nitrocellulose membrane 10 .
[0023] At the other end of the PVC rubber sheet 9, a water-absorbing pad 11 is adhered. ...
Embodiment 2
[0025] The preparation of the cyproheptadine hydrochloride colloidal gold detection card is specifically realized by the following steps:
[0026] Preparation of colloidal gold solution: Take a 1L Erlenmeyer flask, add 495mL of ultrapure water, and then add 1% chloroauric acid (HAuCl 4 ·3H 2 (0) 5mL, be prepared into 500mL0.01% chloroauric acid aqueous solution, add 1% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) solution 5-7mL, continue to stir and heat, when the color of the solution turns into transparent purple red completely, stop heating after maintaining 5min, add water to the original volume, cool to room temperature, and store at 2-8°C for later use;
[0027] Antibody pretreatment: Centrifuge the cyproheptadine hydrochloride antibody to be labeled at 1000r / min, 4°C for 20min, take the supernatant, and dilute to 1mg / mL with 0.01mol / LPBS; or dilute to 1mg with 0.01mol / LPBS / ml, pass through a 0.22μm filter membrane;
[0028] Preparation of colloidal gold markers: ...
Embodiment 3
[0034] The minimum detection limit test of cyproheptadine hydrochloride
[0035] Accurately weigh 0.500g of cyproheptadine hydrochloride antigen, dissolve it in ultrapure water and dilute to 100mL, marked as A 1 solution, take A 1 Add 1mL of the solution to the volumetric flask and add ultrapure water to 1000mL to form A 2 solution, take A 2 Add 1mL of the solution to the volumetric flask and add ultrapure water to 1000mL to form A 3 solution, the concentration of A3 solution is 5ng / mL (5ppb). take A 3 Put 1mL of the solution in 4 test tubes, and label them 1~4 respectively, add 9mL, 4mL, 1mL, 0mL of ultrapure water to the test tubes respectively, and prepare the cyproheptadine hydrochloride antigen solution with concentrations of 0.5ng / mL, 1ng / mL, 2.5ng / mL, 5ng / mL; take another empty test tube, add 5mL ultrapure water, and mark it as No. 5 test tube. Take the liquid in the above-mentioned No. 1-5 test tube as a sample and add it dropwise to the cyproheptadine hydrochlori...
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