2X General Assay Diluent is a mammalian protein-based ELISA additive used to equalize the sample and standard matrices for a more accurate result. Large differences in matrix complexity environment between standard curve and sample wells are the main cause for target analyte under-recovery problems in sandwich ELISA analysis. General Assay Diluent helps equalize matrix complexity disparity issues, reduces non-specific interactions between sample proteins and the plate, and reduces interference from complement and thrombin present in serum and plasma samples. Our 2X concentrated version is helpful for shipping purposes and produces less waste.
General Assay Diluent has been formulated for testing serum, plasma, urine, and cell culture samples in all sandwich ELISA formats. When diluted to its 1X working concentration, it can be used to help minimize matrix complexity differences between the sample (e.g., serum, etc.) and the diluent used to generate the standard curve of the ELISA. Large differences between the sample and the standard diluent matrices will result in under-recovery of the target analyte present in sample wells. Complex and concentrated protein environments (the matrix) present in serum or plasma samples will greatly reduce the antigen-binding efficiency of the plate adsorbed antibodies, resulting in a gross underestimation of the amount of target analyte present in the test samples. General Assay Diluent helps equalize the antibody-binding efficiencies between the standard curve and the sample wells. Mammalian protein additives included in the formulation serve to reduce non-specific interactions between the sample matrix proteins and the plate surface, thereby minimizing background noise. General Assay Diluent also inhibits complement and thrombin activity present in serum and plasma samples. Incorporation of an antimicrobial agent allows for room temperature bench-top use and extensive storage stability. General Assay Diluent, 2X is provided as a 2X concentrate. In order to accommodate increased component quantities present in General Assay Diluent, 2X, the pH has been adjusted to 8.5-8.6, compared to 7.2-7.6 in the 1X formulation. Bulk volumes and custom packaging are available upon request.
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Expires two years from date of manufacture
- Dilute the standard curve, controls, and the samples as necessary. ICT offers several formulations of sample diluents in which to prepare the samples.
- Prepare 1X working strength assay diluent by diluting 1:2 in diH2O. Add 1 part General Assay Diluent, 2X to 1 part diH2O. Mix well.
- Pipette 50-100 μL General Assay Diluent per well into every well of the plate.
- Pipette 50-200 μL of each standard, control, and sample into the plate.
- Run the assay according to the specific ELISA protocol.
- Analyze the data. Because all of the wells, including the standards and controls, received the same volume of assay diluent, there is no need to account for this dilution when calculating the results.