Convenient analysis of protein modification by chemical blotting with fluorogenic “click” reagents

Abstract

Direct visualization of bioorthogonal alkyne or azide handles using fluorogenic azide–alkyne cycloaddition conducted on the surface of a blot membrane. The method eliminates the need for separation steps to remove excess small molecule reagents before attachment of antigen molecules or other visualization handles, and is especially useful for the analysis of peptides and small proteins. A variety of potential fluorogenic reagents are assessed, and sensitivity (<0.1 picomole) similar to current commercially available fluorescence imaging methods is possible.