Protein mapping in circulating leucocytes of breast cancer patients; first-dimensional separation was performed in immobilised pH gradient (IPG) strips (Bio-Rad, USA) in the range of IP 4–7. Following first-dimensional separation, the extruded IPG-strips were equilibrated in gel equilibration buffer I (50 mM Tris–HCl, 6 M urea, 30% glycerol, 2% SDS, 1% DTT), followed by equilibration in buffer II (50 mM Tris–HCl, 6 M urea, 30% glycerol, 2% SDS and 260 mM iodacetamide) for 10 min before loading them onto polyacrylamide gels (12% SDS-PAGE) for the second-dimensional resolution in Mini-PROTEAN 3 (Bio-Rad). Altogether, 74 proteins were consequently identified by MALDI-TOF analysis; data taken from .
Golubnitschaja et al. EPMA Journal 2013 4:6 doi:10.1186/1878-5085-4-6