Antigen-Down Assay Diluent is formulated for use with serum and plasma samples tested in antigen-down ELISA formats. Proprietary additives have been included to reduce the level of nonspecific IgG adsorption to antigen-coated and blocked ELISA well surfaces, as well as enhancing the specific anti-analyte/antigen-antibody signal without denaturing the plate adsorbed antigen molecules.
ICT’s Assay Diluents are proprietary ELISA development additives designed to optimize antigen-down ELISA performance by equalizing 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. Assay Diluents are intended to be added to each well of the plate (~100uL/well) just before the addition of the test samples.
Antigen-Down Assay Diluent is formulated for use with serum and plasma samples tested in antigen-down ELISA formats. Proprietary additives have been included in Antigen-Down Assay Diluent to reduce the level of nonspecific IgG adsorption to antigen-coated and blocked ELISA well surfaces, as well as enhancing the specific anti-analyte/antigen antibody signal without denaturing the plate adsorbed antigen molecules.
A mammalian protein-based buffer and a proprietary detergent additive are incorporated in the antigen-down formulation to minimize the nonspecific background signal that often occurs when analyzing human, porcine, or bovine serum samples. Antigen-Down Assay Diluent also contains the highest molar concentration of calcium chelating agents of all ICT assay diluent product formulations.
Formulated to inhibit interference from complement and thrombin present in serum and plasma samples, Antigen-Down Assay Diluent can also be used as a Sample Diluent for plasma sample analysis in sandwich ELISA configurations. An antimicrobial agent is included in the formulation to allow for bench-top use and storage stability.
Bulk volumes and custom packaging are available upon request.
Once the capture antigen has been coated onto the plate and the plate has been properly blocked with one of ICT’s stabilizing blocker products, pipette 50 – 100 µL of Antigen-Down Assay Diluent into each of the wells of the ELISA plate using a multi-channel pipettor. The Assay Diluent is always added prior to adding the test and control serum/plasma samples to plate wells.
The control and test serum/plasma samples should be diluted to the proper well concentrations using one of ICT’s three different Sample Diluent products. Using an ICT Sample Diluent to dilute the serum and plasma samples will also serve to help minimize the degree of nonspecific IgG binding to the antigen coated and blocked ELISA plate wells. The test (sera and plasma) samples, diluted typically to 1:40 or greater in a sample diluent, should be incubated in the antigen coated ELISA plates for 30 – 60 minutes at room temperature or 37°C.
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