Colloidal gold detection device mixed with spearmint in mint and its decoction pieces, preparation method and application
A detection device and colloidal gold technology, which are applied to measurement devices, biological tests, material inspection products, etc., can solve the problem of inability to quickly detect on-site, and achieve the effects of economy, speed, convenience and low cost.
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Embodiment 1
[0039] Example 1 Preparation of carvone hapten
[0040] Add 1.5g of carvone, 1.4g of carboxymethoxylamine hemihydrochloride, 8mL of methanol, 2mL of pyridine, and 2mL of distilled water into a 100mL three-necked flask, and react at 65°C for 5h. After rotary evaporation, add water, adjust the solution to slightly acidic with dilute hydrochloric acid, extract with ethyl acetate 2-3 times, combine the organic phases, evaporate to dryness, mix the samples, and purify by column to obtain the carvone hapten.
Embodiment 2
[0041] Example 2 Synthesis of Carvone Immune Antigen
[0042] Immunizing antigens were prepared using the carvone hapten. Take carvone hapten 0.1mmol dissolved in 2mL DMF, stir and add 0.2mmol DCC and 0.15mmol NHS. Magnetic stirring was carried out overnight at 4°C. After centrifugation, the supernatant was liquid A. Weighed 140 mg of hemocyanin (KLH) and dissolved it in 10 mL of PBS (pH 8.0) with a concentration of 0.1 mol / L. Add DMF1mL, stir and dissolve to prepare liquid B, under magnetic stirring, gradually drop liquid A into liquid B, and react at 4°C for 12h. After centrifugation, the supernatant was taken, dialyzed with normal saline at 4°C for 3 days, and the dialysate was changed 3 times a day. The obtained whole antigen was dispensed into 0.5 mL centrifuge tubes at a concentration of 1 mg / mL. Store frozen in a -20°C freezer.
Embodiment 3
[0043] Example 3 Preparation of Carvone Monoclonal Antibody
[0044] The monoclonal antibody was prepared by immunizing the antigen with carvone. Use carvone to immunize the antigen and immunize four 6-week-old BALB / C mice after identification. After boosting the immunization three times, blood is collected to measure the titer. When the serum titer no longer rises, the mouse is immunized with twice the dose of antigen without adjuvant. Mice were killed by dislocation of the neck three days later, the spleen was taken under aseptic conditions to prepare splenocytes, mixed with vigorously growing mouse myeloma cells in a 50mL centrifuge tube at a ratio of 8:1, and 30mL of serum-free IPMI1640 medium was added. Centrifuge at 1100r / min for 5min, discard the supernatant, loosen the cell mass, and place in a 37°C water bath. Slowly add 1 mL of 50% PEG-4000 to the cells, drop it within 1 min, and gently stir the sediment at the bottom. After standing for 1 min, slowly and uniformly ...
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