InHuman Cell ELISA KitIt is important to choose various experimental conditions, including:

　　

1. Selection of solid carriers: Many substances can be used as solid carriers, such as polyvinyl chloride, polystyrene, polyacrylamide, cellulose, etc. The form can be concave plates, test tubes, beads, etc. The currently commonly used is 40 hole polystyrene concave plate. Regardless of the type of carrier, screening can be performed before use: coat an equal amount of antigen, react under the same experimental conditions, observe whether the color reaction is uniform, and determine whether its adsorption performance is good.

　　

2. Selection of coated antibodies (or antigens): When the antibodies (or antigens) are adsorbed on the surface of a solid carrier, good purity is required, generally requiring a pH between 9.0 and 9.6. The adsorption temperature, time, and amount of protein also have a certain impact. Usually used at 4 ℃ for 18-24 hours. It is necessary to titrate the appropriate protein coating concentration: different protein concentrations (0.1, 1.0, and 10 μ g/ml, etc.), and observe the OD value of the positive sample under the same experimental conditions. Choose the concentration with the highest OD value and the lowest protein content. For most proteins, it is usually 1-10 μ g/ml.

　　

3. Selection of working concentration for enzyme-linked immunosorbent assay (ELISA): First, use the direct human cell ELISA kit ELISA method to titrate the initial potency (see section on enzyme-linked immunosorbent assay). Then fix other conditions or use the "matrix method" (coating, reference sample for the test sample, and enzyme-linked antibody at different dilutions) to accurately titrate the working concentration in the formal experimental system.

　　

4. Selection of enzyme substrates and hydrogen donors: The selection of hydrogen donors requires low cost, safety, obvious color reaction, and colorless. Some hydrogen donors (such as OPD) have potential carcinogenic effects and should be protected. If conditions permit, non carcinogenic and highly sensitive hydrogen donors such as TMB and ABTS should be used, which are currently satisfactory hydrogen donors. After a period of substrate action, strong acid or strong base should be added to terminate the reaction. The general substrate action time should be 10-30 minutes. The substrate solution must be freshly prepared, especially with the addition of H2O2 before use.