Colloidal gold test strip for detecting neomycin residual and use method and application thereof
A technology of colloidal gold test paper and neomycin, which is applied in the field of veterinary drug residue analysis and immunology, and can solve problems such as neomycin immunochromatographic test strips that have not yet been seen.
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Embodiment 1
[0024] Embodiment 1 Preparation of colloidal gold test strips
[0025] 1.1 Preparation and purification of monoclonal antibodies
[0026] 1.1.1 Preparation of ascites: several Balb / c mice were taken 7 days before inoculation, and each mouse was pretreated by intraperitoneal injection of 0.5ml Freund's incomplete adjuvant. Use RPMI 1640 basal medium to suspend the cells expanded and cultured by the hybridoma cell EDC / 5G04 with the preservation number CCTCC NO:C201144, and adjust the number of cells to 1×10 6 cells / mL, each mouse was inoculated with 0.5ml intraperitoneally. Ascites was collected when the abdomen of the mice was obviously enlarged, the spirit became poor, and they were immobile when they were dying.
[0027] 1.1.2 Purification of monoclonal antibody: refer to the method in Zhu Liping's "Common Experimental Methods in Immunology" People's Military Medical Publishing House 2000 Edition: Take 5 ml of the obtained mouse ascites and mix it with an appropriate amount...
Embodiment 2
[0047] The using method and application of embodiment 2 colloidal gold test strip
[0048] 2.1 Optimization of colloidal gold test strip detection method
[0049] When the coating concentration is 0.005mg / ml, the negative blank test line has no obvious color development, indicating that the concentration of the original coating is too low, and the amount of intercepting colloidal gold-labeled antibody is too small, and the concentration of the original coating needs to be increased. When the coating concentration is When the concentration is greater than 0.01mg / ml, the detection sensitivity cannot meet the requirements. When the coating concentration is 0.01mg / ml, the negative blank has a clear color and can detect the standard substance of 50μg / L, and finally determine the neomycin coating. The coating concentration is 0.01mg / ml.
[0050] After determining the original concentration of the coating, coat different concentrations of goat or rabbit anti-mouse IgG, so that the c...
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