Immunochromatography time resolution fluorescence kit for synchronization detection of mixed pollution of fumonisin B1 and other four fungaltoxin and application thereof
A time-resolved fluorescence and fumonisin technology, applied in fluorescence/phosphorescence, measuring devices, analytical materials, etc., can solve problems such as low sensitivity, high incidence of mycotoxins, and inability to perform quantitative detection, achieving good specificity and sensitivity high effect
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Embodiment 1
[0039] Example 1 Acquisition of aflatoxin B1 monoclonal antibody
[0040] The aflatoxin B1 monoclonal antibody is secreted and produced by the hybridoma cell line 3G1 with the deposit number CCTCC NO.C201014, and is specifically prepared in advance according to the method reported in the patent with the authorization number ZL201210117614.9. The preparation method is: the obtained hybridoma The tumor cell line 3G1 was injected into BALB / c mice pretreated with incomplete Freund's adjuvant, and the ascites of the mice was collected and purified to obtain aflatoxin B1 monoclonal antibody. Among them, the purification method is octanoic acid-ammonium sulfate method, and the specific operation is as follows: filter the mouse ascites with double-layer filter paper, centrifuge the filtered ascites at 4°C for more than 15 minutes at 12000 r / min, absorb the supernatant, mix the supernatant with 4 times the volume Acetate buffer solution was mixed, and n-octanoic acid was slowly added w...
Embodiment 2
[0042] Example 2 Acquisition of Ochratoxin A Monoclonal Antibody;
[0043] The ochratoxin A monoclonal antibody is secreted and produced by the hybridoma cell line 1H2 with the deposit number of CCTCC NO.C201329, and is specifically prepared in advance according to the method reported in the patent with the application number of 201310115921.8. The preparation method is: the hybridoma cell line 1H2 It was injected into the abdomen of BALB / c mice previously treated with incomplete Freund's adjuvant, and the ascites of the mice was collected and purified to obtain ochratoxin A monoclonal antibody. The purification method is octanoic acid-ammonium sulfate method, and the specific steps are: filtering mouse ascites fluid with double-layer filter paper, centrifuging at 12000 r / min for more than 15 min at 4°C, sucking the supernatant, and mixing the obtained ascites supernatant with 4 times the volume of vinegar. Mix with salt buffer, slowly add n-octanoic acid with stirring, the vo...
Embodiment 3
[0045] The acquisition of embodiment 3 zearalenone monoclonal antibody
[0046] The zearalenone monoclonal antibody is secreted and produced by the hybridoma cell line 2D3 with the deposit number CCTCC NO.C201328, and is specifically prepared in advance according to the method reported in the patent with the application number 201310115825.3. The preparation method is: the hybridoma cell line 2D3 was injected into the abdomen of a BALB / c mouse pretreated with incomplete Freund's adjuvant, and the ascites of the mouse was collected and purified to obtain a zearalenone monoclonal antibody;
[0047] The purification method is octanoic acid-ammonium sulfate method, and the specific steps are: filtering mouse ascites fluid with double-layer filter paper, centrifuging at 12000 r / min for more than 15 min at 4°C, sucking the supernatant, and mixing the obtained ascites supernatant with 4 times the volume of vinegar. Mix with salt buffer, slowly add n-octanoic acid with stirring, the v...
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