Quantitation

The NanoPro 1000's high-resolution IEF capillary separation provides outstanding resolution of phosphoprotein isoforms. By using a pan-specific antibody to recognize all non-phosphorylated and phosphorylated isoforms, relative phosphorylation levels can be estimated. In the example shown below, MCF10A breast cancer cells were serum starved (control) and then stimulated with 10 ng/mL EGF for 25 minutes (stimulated). Simple area-under-the-curve calculations show that the proportion of phosphorylated MEK1 increased from 70% to 92% after stimulation.