Commonly used in immunoassay enzyme enzyme-linked immunoassay enzyme technology with high sensitivity is the key to select enzymes with biological amplification as a marker. The enzyme used in the enzyme immunoassay must meet the following conditions: 1 The purity of the enzyme is high, the conversion rate of the catalytic reaction is high, and the specificity of the enzyme action is strong; After chemical coupling with antigen and antibody protein molecules, it still maintains high catalytic activity; 3 stability in use and storage stability; 4 method for measuring enzyme activity is simple, sensitive and fast; 5 in body fluid to be tested It is best not to have the same enzyme as the labeled enzyme to prevent interference; 6 There should be no substrate, reaction inhibitor and other interference factors in the solution to be tested; 7 The source, purification and supply of the enzyme are more convenient, 8 homogeneous The enzyme used in the enzyme immunoassay also requires that the activity of the enzyme shows inhibition or activation when the antibody is combined with the hapten-enzyme conjugate. The most commonly used tracer enzymes in the experiment are horseradish peroxidase, alkaline phosphatase, glucose oxidase, β-D-galactosidase and so on. Among them, according to the size of the molecular weight and the characteristics of enzyme reactivity, it should be selected in the solid-phase or non-solid-phase enzyme-labeled immunoassay. In the enzyme-linked immunoassay technique, since the enzyme-labeled complex often enters the cell through the cell membrane and binds to the target molecule, the label used should select an enzyme with a low molecular weight and high enzyme activity. In the quality evaluation of labeled enzymes, two indicators are usually needed: one is the RZ ratio and the other is the activity unit. RZ represents the purity of the enzyme, that is, the ratio of the maximum absorbance of the active part to the inactive part in the enzyme protein. The physicochemical properties of an enzyme are usually used to determine whether it is suitable as a marker. Compared with anisotropic markers, enzymes have a variety of measurement methods, and the enzymes with small molecular weights can also be determined by second-stage amplification, which shows that enzymes are a general-purpose marker and can be widely used in various analyses. From simple operations to highly sensitive automated procedures, enzymes are an excellent marker, but enzymes are more susceptible to environmental factors and inactivation than fluorescent or chemiluminescent compounds. In addition, the limitation of the enzyme immunoassay method in terms of low detection limit or detection range needs to be further improved, but this limitation can be eliminated by selecting an endpoint assay.

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