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.
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General Assay Diluent – Mammalian proteins
IgM-Reducing Assay Diluent – Mammalian proteins
Neptune Assay Diluent – Non-mammalian proteins
Antigen-Down Assay Diluent – Mammalian proteins
United States
Expires 18 months - two years from date of manufacture
- To quickly determine which Assay Diluent formulation is best for the assay, all four Assay Diluents can be run in parallel on the same IgG-coated ELISA plate.
- Determine the initial dynamic analyte detection range of the ELISA curves that were prepared in each of the four different Assay Diluent types.
- Make test spikes into serum samples.
- Read the spiked serum sample wells from each of the four AD-associated standard curves to see which Assay Diluent gives the best approximation of the spike concentration that was added to the serum samples.
- In cases where a high level of matrix complexity interference is not detected, an additional Assay Diluent may not be required to obtain a useful standard curve and derived sample analysis.