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1 Avoid contamination so be very careful that you avoid spillover from adjacent wells. 2 Don't allow the plate to dry out. 3 Make sure that all incubations are carried out in 100% humid conditions. (wrapping the covered plate in a wet paper towel and then placing in a sealed bag works) 3 Wash the plate well and carefully with PBS-tween between steps at least 3 times. 4 Use fresh buffers all at the correct pH. 5 Don't use any azide in your buffers if you use peroxidase conjugates. There are more but it takes experience to do an ELISA test well.
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