Heavy metal enzyme label and application thereof
A technology of heavy metals and enzyme labels, applied in the direction of microorganisms, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of unavailable rapid detection, high detection cost, and high cost, and achieve HRP activity loss, low synthesis cost, The effect of simple preparation method
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Embodiment 1
[0056] Example 1. A kit for detecting whether the sample contains heavy metal copper
[0057] 1. Composition of the kit
[0058] (1) Composition of the kit of the present invention
[0059] 1. Original coating: anti-copper monoclonal antibody, which is the monoclonal antibody secreted by the hybridoma cell line Cu-EDTA 6A9 with deposit number CGMCC No.3987;
[0060] 2. Enzyme-labeled substance: The enzyme-labeled heavy metal copper is a cross-linking product formed by a complex and horseradish peroxidase through covalent bonds; the complex is a combination of p-aminobenzylethylenediaminetetraacetic acid and BSA. The conjugate formed by covalent bond is connected with the heavy metal copper through coordination bond to form a complex.
[0061] 3. Coating buffer: 0.05M, pH9.6 carbonate buffer;
[0062] 4. Washing solution: Each 1 liter washing solution is prepared as follows: 8.0g NaCl, 0.2g KH 2 PO 4 , 2.96gNa 2 HPO 4 ·12H 2 O. 1 mL of Tween-20 was dissolved in water an...
Embodiment 2
[0118] Example 2. Application of the kit
[0119] The EDTA-Cu chelate standard solution prepared above was diluted with the sample diluent to the following different Cu ion concentrations: 10ng / mL, 5ng / mL, 2.5ng / mL, 1.25ng / mL, 0.625ng / mL, 0.31ng / mL, 0.156ng / mL, 0.078ng / mL and 0.039ng / mL.
[0120] (1) Antibody coating: 1 mg / mL monoclonal antibody was diluted at 1:2000 and added to the ELISA plate, 100 μL per well, incubated at 37°C for 3 hours; poured out the solution in the ELISA plate, washed with Wash the plate 4 times with liquid and spin dry;
[0121] (2) Add the above EDTA-Cu chelate standard solution (experimental well) with different concentrations to the ELISA plate in step (1), 50 μL per well, and no EDTA-Cu chelate standard solution is added to the control well solution and add 50 μL of sample diluent;
[0122] (3) Add 0.5 μg / mL enzyme labeling substance to the above experimental wells and control wells, 50 μL per well; incubate at 37°C for 30 min; pour off the so...
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