In enzyme-linked solid-phase immunoassays, especially in the indirect ELISA method, the problem of negative background is often encountered. The main reason is that the background is too high and the background value of different specimens is quite different. Background or background is the non-specific color development in ELISA. The color of the substrate without enzyme action is called blank. If the substrate solution is not properly prepared, a certain blank value will appear. As long as this blank value is not too high (A <0.05), it can be subtracted from each measured value and does not affect the experimental results. What is discussed here is the background of negative specimens that do not contain the test antigen or antibody in ELISA. In theory, only negative specimens can eventually cause color reaction, so why does the background appear? Adsorption of solid-phase carriers: In enzyme immunoassays, ELISA is characterized by the use of means to separate free and bound enzyme conjugates. The solid-phase carrier and the antigen or antibody adsorbed thereon form a solid-phase antigen or solid-phase antibody, that is, an immunosorbent (Immu-nosorbent). The commonly used solid-phase carrier polystyrene has mainly hydrophobic groups on its surface and binds to proteins through hydrophobic bonds. This physical adsorption is non-specific. Although the molecular weight, isoelectric point, and concentration of the protein have a certain effect on the amount of adsorption, in general, various proteins can be adsorbed on the surface of the polystyrene carrier. Therefore, in addition to purposefully adsorbing the antigen or antibody on the carrier during coating, the adsorption of interfering substances may occur in each step of the ELISA, resulting in coloration that is ultimately unrelated to the antigen-antibody reaction. This is The main reason for forming the background. Liyang Paper Products Co., Ltd. was established in 1999,a professional enterprise integrated in R&D,Marketing and Manufacturing.Liyang Printing provide best solutions and service to various fields of industry.For example,we can make nice and luxury Softcover Book Printing.Such as Softcover Book,Softcover Book Printing ,Softcover Children Book,Softcover Book Handmade,etc.Our softcover Book with high quality are exquisite printing for your products.
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Causes and elimination methods of negative background in ELISA