How many proteins can be identified from a single gel band?

This depends on the complexity of the sample that was loaded on the gel.  From our experience we have identified as many as 70 proteins from a single gel band when complex cell lysate was ran on 12%T minigel.  We strongly recommend not to overload the gel as it will affect the resolution and separation of proteins. Moreover, it can result in “carry-over” of abundant peptides in LC resulting in identification of same protein repeatedly in different samples. 

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