The effect of staining on the immunoreactivity of nitrocellulose bound proteins.

It was found that Coomassie blue staining of proteins electroblotted onto nitrocellulose resulted in a significant decrease in subsequent immunoreactivity, relative to unstained proteins. This was not uniform, and resulted in distortion of the pattern detected by immunoreaction as well as an overall decrease in sensitivity. Staining with amido black resulted in no such alteration. Additionally, it was observed that amido black is completely leached from the nitrocellulose strip during immunoreaction, whereas no leaching is apparent with Coomassie blue. This may contribute to the observed effect of staining on immunoreactivity.