General Assay Diluent, 2X

Milestones Common to All Sandwich ELISA Development Projects, 1. Acquisition of adequate quantities of quality antibody (IgG) and analyte components necessary to initiate project., 2. Preparation or acquisition of the sandwich IgG-HRP conjugates., 3. Demonstration of basic assay feasibility (dose response)., 4. Optimization of capture antibody coating., 5. Resolution of sample matrix interference issues., 6. Final optimization of sandwich IgG-HRP conjugate concentration, considering the inhibitory effects of the sample matrix., 7. Demonstration of ELISA performance capabilities via assessment of assay linear dynamic range, sensitivity, linearity, standard recovery efficiency, and precision., , For detailed protocols and information on each of these ELISA development milestones, please see the Antibody-Sandwich ELISA Development Kit Manual.

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.