"The Caffeine ELISA is an immunoassay for the quantitative and sensitive detection of Caffeine. This test is suitable for the quantitative and/or qualitative screening of Caffeine in water, urine, and saliva samples. If necessary, positive samples can be confirmed by HPLC, GC/MS, or other conventional methods. Note: This assay is intended For Research Use Only. The test is a direct competitive ELISA which is based on the recognition of Caffeine by specific antibodies.
Caffeine, when present in a sample, and a Caffeine-HRP analogue compete for the binding sites of mouse anti-Caffeine antibodies in solution.
The Caffeine antibodies are then bound by a second antibody (goat anti-mouse) immobilized on the wells of the microtiter plate. After a washing step and addition of the substrate solution, a color signal is generated. The intensity of the blue color is inversely proportional to the concentration of Caffeine present in the sample.The color reaction is stopped after a specified time and the color is evaluated using an ELISA reader. The concentrations of the samples are determined by interpolation using the standard curve constructed with each run."