Indirectly racing ELISA detecting method for cadmium ion
A detection method, the technology of cadmium ions, applied in the biological field, can solve the problems of insufficient reporting and the inability to repeatedly synthesize artificial antigens, and achieve the effect of guaranteeing interests, low cost, and sensitive detection
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[0034] Example 1
[0035] The indirect competitive ELISA method for detecting cadmium ions by using anti-cadmium ion monoclonal antibody includes the following steps:
[0036] (A) Cadmium ion coupling to hemocyanin (KLH) with iEDTA (i.e. 1-(4-isothiocyanatobenzyl)ethylenediamine-N,N,N',N'-tetraacetic acid), BALB / c mice were immunized with incomplete Freund's adjuvant, spleen cells of the immunized mice were fused with SP2 / 0 myeloma cells, and positive cell lines that stably secreted anti-cadmium ion monoclonal antibody (MAb-Cd) were screened and expanded. Culture, inject cells into mice to induce ascites, and purify to obtain anti-cadmium ion monoclonal antibody.
[0037] (B) The detection antigen Cd-iEDTA-BSA was obtained by coupling cadmium ions to the carrier protein BSA with iEDTA.
[0038] (C) Preparation of antigen pre-coated strips:
[0039] (1) Coating: Dilute Cd-iEDTA-BSA to 0.2 μg / mL with coating buffer (0.05M sodium carbonate buffer, pH 9.6), and add 100 μL per m...
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[0060] Example 2
[0061] The indirect competitive ELISA method for detecting cadmium ions by using anti-cadmium ion monoclonal antibody includes the following steps:
[0062] (A) Using iEDTA to couple cadmium ions to limpet hemocyanin (KLH), BALB / c mice were immunized with incomplete Freund's adjuvant, and spleen cells of the immunized mice were fused with SP2 / 0 myeloma cells. The positive cell line that stably secretes anti-cadmium ion monoclonal antibody (MAb-Cd) is expanded and cultured, and the cells are injected into mice to induce ascites, and the anti-cadmium ion monoclonal antibody is purified and obtained.
[0063] (B) The detection antigen Cd-iEDTA-BSA was obtained by coupling cadmium ions to the carrier protein BSA with iEDTA.
[0064] (C) Preparation of antigen pre-coated strips:
[0065] (1) Coating: Cd-iEDTA-BSA was diluted to 0.2 μg / mL with coating buffer (0.05M sodium carbonate buffer, pH 9.6), and 100 μL per microwell was added to the micron of polystyrene ...
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