Adapting the Single-Cell Western Protocol to Detect Histone Modifications

The Single-Cell Western^TM protocol is highly versatile and can easily be adapted for different biological applications and protein targets. Researchers studying signaling proteins or other targets that require treatment of cells prior to analysis can add a drug, cytokine, or other form of pre-treatment directly to scWest chips after cells have been captured and before running the chip on Milo. Cells can also be treated just prior to settling them onto scWest chips. As a result, the flexible Single-Cell Western workflow allows time-dependent experimental manipulations to be easily performed before lysis and electrophoresis. Here we describe a two-step, on-chip pre-treatment protocol to measure heterogeneity of modified histones - a class of proteins that is challenging to measure but critical in regulating gene expression. This new measurement capability could be key to unlocking new discoveries in the field of epigenetics.