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How to choose the right microplate?

Posted by Admin | 31 Jan

1: According to the bottom of the microplate can be divided into flat bottom, U-type bottom, and V-type bottom The refractive index of the flat base is low, suitable for detection in the microplate; the U microplate refractive index is convenient for sampling, sampling, and mixing, can directly observe the color change without placing on the microplate, so as to determine whether there is a corresponding immune reaction. The microplate of the V base can accurately absorb the sample.

 

2: According to the different binding abilities of the microplate and protein and other molecules, it is divided into high binding force, medium binding force and amination High binding force: after the surface treatment, the protein binding ability is greatly enhanced, reaching 300 ~ 400 ng IgG / cm2, and the molecular weight of the main bound protein is> 10 kD. The use of this class of microplate can improve the sensitivity, and can reduce the concentration and amount of coated protein relatively, which is easier to produce non-specific reactions. After coating with antigen or antibody, the non-ionic detergent cannot effectively seal the site of the unbound protein, and the protein should be used as the sealant agent.

 

Medium binding force: passive binding to protein by surface hydrophobic bond, suitable as solid phase carrier of macromolecular protein with molecular weight> 20 kD, with protein binding capacity of 200~300ng IgG / cm2. Due to the characteristics of its only binding to macromolecules, it is suitable for solid-phase carriers as unpurified antibodies or antigens to reduce the potential for nonspecific cross-reactivity. The plate can be an inert protein or a nonionic detergent as a sealing solution.

 

Aminidination: the surface modification has a positively charged amino group, whose hydrophobic bond is replaced by a hydrophilic bond. This class of microplate is suitable as a solid-phase carrier for small-molecule proteins. Using a suitable buffer and pH, the surface binds to negatively charged small molecules via ionic bonds. Due to the hydrophilic properties of its surface and its ability to be covalently bound by other cross-linkers, it can be used to fix protein molecules soluble in decontamination agents such as Triton-100, Tween 20, etc. The defect of this plate is because of reduced hydrophobicity; moreover, the surface needs to be effectively closed. Because of the hydrophilic and covalent surface properties, the sealing solution used must be able to interact with any functional group in the nonreactive amino group and the selected crosslinker.

 

3: According to the color is divided into transparent, black, the white Transparent is the most commonly used for the most general enzyme-linked immunization experiments. Relative to the transparent board, there are also opaque boards used for luminous detection, generally black and white. The black microplate itself has light absorption, so its signal is much lower than the white plate, so it is generally used to detect strong light, such as fluorescence detection. The white microplate is used for weak light detection, often used for general chemiluminescence. In addition, the black microplate can also weaken the problem of non-specific reactions. Some customers will ask, with the general microplate can be luminous detection, the answer is not possible, because the light emitted from the chemiluminescence reaction is isotropic, that is to say, the same emission in all directions. If a transparent plate is used, the light not only diverges from the vertical direction, but also from the horizontal direction. It easily passes through the gaps between the holes and the hole walls. In this way, the effect between adjacent pores will be large when the light is strong, so the chemiluminescence cannot be tested with a transparent microplate.

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