Quantitative enzyme-linked immunoabsorbent assay method based on moving reaction boundary electrophoresis titration
A technology of mobile reaction and detection method, which is applied in the direction of chemical reaction of materials and material analysis by observing the influence on chemical indicators, so as to reduce costs and simplify ELISA detection.
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[0024] Example 1
[0025] Such as Figure 1~3 As shown, this embodiment uses ELISA-MRB-ET to detect standard samples of SEB enterotoxin with different concentrations and known under the same conditions, and establishes the functional relationship between MRB movement distance and substrate concentration, along the electrophoresis channel Mark the concentration scale; in the sample detection, use the same conditions for ELISA-MRB-ET detection, and the concentration scale corresponding to the stop position of the formed MRB is the concentration of the substrate in the sample; specifically includes the following steps:
[0026] Step 1. Fill the gel in the electrophoresis channel and coat and block the primary antibody.
[0027] The electrophoresis channel is a straight electrophoresis channel, and its length, width, and height are respectively 20mm, 1.0mm and 0.5mm.
[0028] Both ends of the electrophoresis channel are respectively provided with a cathode chamber and an anode chamber.
[...
Example Embodiment
[0052] Example 2
[0053] The standard sample in this example is glycosylated hemoglobin, and the HbA1C in human whole blood is determined by the double antibody sandwich method.
[0054] The primary antibody of this example was 10 mg / L, 200 μL of haptoglobin, which was coated on the cathode compartment overnight at 4° C., and washed with phosphate buffer solution 3 times.
[0055] The blocking in this embodiment refers to: using 200 μL of 3% BSA to block the blank sites on the plate, incubating for 1 h at 37° C., washing with phosphate buffer solution 3 times to remove excess liquid.
[0056] The difference compared with Example 1 is that the multiple dilution concentrations of the standard samples of this example are 5μg / L, 2.5μg / L, 1.5μg / L, 0.625μg / L, 0.3125μg / L and 0μg / L respectively. .
[0057] The glycosylated hemoglobin can reflect the patient's blood glucose control status in the past 8-12 weeks, and is the new standard for diabetes diagnosis and the "gold standard" for treatme...
Example Embodiment
[0058] Example 3
[0059] The standard sample of this embodiment is the residual thion on the lettuce.
[0060] Said thion is a broad-spectrum insecticide. my country requires that the maximum residue of agricultural products should be less than 0.1mg / kg, and it cannot be detected on vegetables and fruits.
[0061] The primary antibody in this example is a goat anti-rabbit antibody, diluted by a factor of 2000, coated the cathode compartment overnight at 200 μL and 4° C., and washed with phosphate buffer solution 3 times.
[0062] The difference compared with Example 1 is that the dilution concentrations of the thion standard sample in this example are 500 ng / mL, 100 ng / mL, 25 ng / mL, 12.5 ng / mL, 6.5 ng / mL and 0 ng / mL, respectively.
[0063] The experimental conditions of the enzyme-linked immunosorbent assay in this embodiment are: 10 μL of standard sample, 45 mL of enzyme label and 45 mL of antibody are incubated in the cathode chamber at 37° C. for 30 min, and washed with phosphoric a...
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