Assay Diluent Optimization Pack

1. To quickly determine which Sample Diluent formulation is best for the assay, all three Sample Diluents can be run in parallel on the same IgG or antigen-coated ELISA plate., 2. Determine the initial dynamic analyte range of the ELISA curves that were prepared in each of the three different Sample Diluent types., 3. Make test spikes into serum samples., 4. Read the spiked serum sample Wells on each of the three SD-associated standard curves to see which Sample Diluent gives the best approximation of the spike concentration that was added to the serum samples., 5. In cases where a high level of matrix complexity interference is detected, inclusion of an additional Assay Diluent reagent may also be required to bring the calibrator Well antibody-to-antigen binding kinetics closer to the sample Well binding kinetics.

Achieving a high signal-to-noise ratio and a solid standard curve are goals of any ELISA development project, yet a number of complex issues may arise, presenting challenges to assay optimization. Many issues stem from differences between the sample and calibrator matrices and nonspecific binding of immunoassay components., , Most of the time invested in developing a new ELISA/EIA/RIA will involve selecting the proper calibrator matrix that matches most closely to the often very complex biological sample matrix that is being evaluated. Assay Diluents are additive components used in an ELISA to equalize any differences between the sample matrices (serum, plasma, urine, cell culture fluid) and the calibrator diluent used to generate the standard curve of the ELISA. ImmunoChemistry Technologies offers four ELISA development buffers that were designed to address these complex matrix effects and other assay concerns., , During ELISA development, it is often helpful to compare a number of different Assay Diluent formulations to better match the matrix complexity present in the sample Wells with the standard curve calibrator matrix Wells. Assay Diluent Optimization Pack, containing four ELISA development buffers (General Assay Diluent, IgM-Reducing Assay Diluent, Neptune Assay Diluent, and Antigen-Down Assay Diluent), offers an economical method of addressing matrix effects and optimizing signal-to-noise ratio in ELISA development projects.