ELISA test box for detecting zearalenone and preparing and detecting method thereof
A single technology of zearalenone and zearalenone is applied in the field of ELISA test box and preparation and detection of zearalenone. Expensive and other issues, to achieve the effect of simple operation, convenient use, and rapid detection
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Embodiment 1
[0020] The coated plate is coated with solid-phase antigen, which can be 96 or 48 or 24-well microplate, with 10mmol / LpH9.5 Na 2 CO 3 -NaHCO 3 Dilute the vomitoxin-ovalbumin (ZEN-OVA) to 1.0 μg / mL, add 100 μl to each well of a 96-, 48-, or 24-well microplate, place at 5°C overnight, discard the coating Wash twice, add phosphate buffer solution containing 5% bovine serum albumin, pH 7.5, block overnight at 8°C, discard the blocking solution, blow dry, seal the strips and store in a freezer at -20°C;
[0021] The standard zearalenone is obtained by diluting the pure zearalenone, the diluent is deionized water, and the concentration of ZEN is: 25ng / mL;
[0022] Preparation of zearalenone-bovine serum albumin (ZEN-BSA) conjugate: Dissolve ZEN in 100mL pyridine, add carboxymethylhydroxylamine hydrochloride, stir overnight at room temperature, blow dry with nitrogen, add 55ml distilled water , use sodium hydroxide solution to adjust the pH to about 9, then extract with dichlorome...
Embodiment 2
[0026] The coated plate is coated with solid-phase antigen, which can be 96 or 48 or 24-well microplate, with 55mmol / L pH9Na 2 CO 3 -NaHCO 3 Dilute the vomitoxin-ovalbumin (ZEN-OVA) to 0.55 μg / mL, add 100 μl to each well of a 96 or 48 or 24-well microplate, place it overnight at 2°C, discard the coating solution, washed 5 times, added phosphate buffer solution containing 1% bovine serum albumin, pH 7, blocked overnight at 5°C, discarded the blocking solution, dried, sealed the strips and stored at -40°C;
[0027] The standard zearalenone is obtained by diluting the pure zearalenone, the diluent is deionized water, and the concentration of ZEN is: 50ng / mL;
[0028] Preparation of zearalenone-bovine serum albumin (ZEN-BSA) conjugate: Dissolve ZEN in 3 mL of pyridine, add carboxymethylhydroxylamine hydrochloride, stir overnight at room temperature, blow dry with nitrogen, add 100 mL of distilled water , adjust the pH to about 8.5 with sodium hydroxide solution, then extract 5 ...
Embodiment 3
[0032] The coated plate is coated with solid-phase antigen, which can be 96 or 48 or 24 well microwell plate, with 100mmol / LpH10 Na 2 CO 3 -NaHCO 3 Dilute the vomitoxin-ovalbumin (ZEN-OVA) to 0.1 μg / mL, add 100 μl to each well of a 96-, 48-, or 24-well microplate, place at 8°C overnight, discard the coating solution, washed 3 times, added phosphate buffer solution containing 3% bovine serum albumin, pH 8, blocked overnight at 2°C, discarded the blocking solution, dried, sealed the strips and stored at -30°C;
[0033] The standard zearalenone is obtained by diluting the pure zearalenone, the diluent is deionized water, and the concentration of ZEN is: 0.001ng / mL;
[0034] Preparation of zearalenone-bovine serum albumin (ZEN-BSA) conjugate: Dissolve ZEN in 51.5mL pyridine, add carboxymethylhydroxylamine hydrochloride, stir overnight at room temperature, blow dry with nitrogen, add 10ml Distilled water, adjust the pH to about 9.5 with sodium hydroxide solution, then extract 4 ...
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