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How to wash the ELISA plate?
Correct washing of the wells is very crucial for a successful ELISA. Deionized or distilled water should be used when diluting the concentrated Wash Buffer.
Using multi-channel pipette
Firstly, check the strips in the plate so that they are firmly in the plate holder. Numbering the strips may be useful in the event that strips become loose while decanting. Secondly, empty the liquid in the plate by decanting. Fill each well with the volume of Wash Buffer specified in the kit manual. Set a timer for the recommended time and let the Wash Buffer soak in the wells. Decant the wells by inverting the plate and rapping it against clean paper toweling. Repeat this process as directed in the kit manual. After the last decanting, remove any remaining Wash Buffer by inverting the plate and blotting it dry by rapping it firmly against clean paper toweling on a hard surface. Do not let the wells to sit dry. Proceed immediately to the next step in the kit manual.
Using manifold dispenser or autowasher
Equip the dispenser or autowasher with an appropriate vacuum supply (refer to manufacturer’s guidelines). Ensure that each cannulae is dispensing and aspirating properly. Set the volume of Wash Buffer in each well and soaking time for each washing. Then put the plate in. Ensure that no Wash Buffer is left in the wells after aspiration. Do not over-aspirate the wells by allowing the aspiration apparatus to sit in the wells after the aspiration.
Washing the plate too rapidly or too slowly, incomplete washing or aspirating, and letting the wells to sit dry are all factors that will affect the precision of the ELISA.