To select the key points in the elisa experiment steps

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Many customers responded that the seemingly simple elisa experiment is a little unreasonable in the operation process, which will cause many problems, affect the detection accuracy and reduce the test quality. Such as flower board, false positive, full color, full color, low color space, etc., which requires us to summarize in the experimental process, gradually improve, and select the key points in the elisa experiment steps. First: the choice of coating solution Carbonate buffer is usually selected 9.6. But sometimes because of the test requirements, the packet buffer solution is a special kind of raw material that can be used in neutral packaging. Attention should be paid to the following principles: Protein and polystyrene solid phase carriers are bound by physical adsorption, depending on the hydrophobic group in the solid phase and the hydrophobic interaction of the surface structure of the protein molecule, the physical adsorption is non-specific, protein Molecular weight, isoelectric point, large concentration, small molecular protein protein usually contains more hydrophobic groups, so it is easier to adsorb on the surface of the solid support. Testing also has a strong theory of practice, and the last one can be applied to our tests. In addition to the commonly used coating solution of the 9. 6 carbonate buffer just mentioned, and phosphate buffer and 7-8 7. 2 - buffer. Second: Closed The following package is followed by a high concentration of protein solution, coating process. Enclosed are immunological rejection and adsorption steps that fill a large number of unrelated proteins with these voids, and interfering substances. Elisa kit commonly used seal: 0. 05% - 0. 5% bovine serum albumin; 10% or 1% gelatin bovine serum; skim milk powder, relatively inexpensive, can be used for high concentrations (5% ~ 10%); Some rare use of various animal serum (mainly to eliminate interference with similar proteins and casein). But what to choose, according to the specific practice of the experiment. Third: washing the board It can be said that in the operation, cleaning is the main key technology. Since the adsorption of proteins by plastics such as polystyrene is ubiquitous, in order to achieve the separation of the free enzyme and the objective label, the removal of free and adsorbed non-specific interfering substances in the pores of the sheet, washing, and Non-specific adsorption of interfering substances washed. Therefore, there will be some error in the washing board, very human factors (of course, there are washing machines except conditions), which are incomplete or string clear holes, the influence of the system is so sensitive, but not small. Fourth: add the antibody (anti-specimen and two) attention to the tip of the gun, the gun head. The sample is diluted with dilution and can also be diluted with a diluted solution. If you want to add two resistors, but also pay attention to the two reverse working concentrations of waste, too high, too low light color.

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