When performing ELISA using Nunc-Immuno MaxiSorp Plates, what are some recommendations for reducing high background readings and non-specific binding?
Assay sensitivity depends strongly on an efficient removal of non-specific reacting molecules. High background readings and coating instability can be eliminated by addition of a blocking step after the first coating. The excess surface is then occupied by indifferent molecules. We recommend washing three times after each coating step by using a solution of 0.15 M phosphate buffer (pH 7.2) with 0.2 M NaCl and 0.05% Tween 20. For blocking, we recommend using 0.5% BSA, 1% casein or 1% gelatin in 0.15 M phosphate buffer (pH 8.2) or carbonate buffer (pH 9.6). See Nunc Bulletins No. 7, 8, and 9.
Related Questions
- When performing ELISA using Nunc-Immuno MaxiSorp Plates, what are some recommendations for reducing high background readings and non-specific binding?
- Using the kit LowCell# ChIP I got very high background; even with SAT2 I got the band performing end-point PCR. What can be the problem?
- Does mathematics teaching in high performing countries resemble the reform movements recommendations?
