Microfluidic devices coupled to MS promises low manufacturing costs, low sample consumption and channels with a high surface area to volume ratio and tailorable functional groups. Here, we describe a thiol-ene-based microfluidic chip that is capable of sample loading, desalting and on-chip reversed phase chromatography prior to on-chip electrospray ionization for the intact analysis of proteins and peptides, as well as, for on-chip bottom-up proteomics workflows coupled directly with mass spectrometry.