An important step in the proteomic analysis is the optimal extraction and the processing of protein material in order to ensure the highest sensitivity in downstream protein detection. Cell lysis is important for proteomic profiling with an eventual aim to discover differently expressed proteins (DEPs). Herein for cell lysis, two sample preparation protocols were compared: 0.2% SDS-based solubilization combined with the 1DE-gel concentration (Protocol 1) and osmotic shock (Protocol 2). HaCaT keratinocytes were cultured with/without sodium dodecyl sulfate (SDS) and used to test the hypothesis that SDS might be associated with the initiation and development of malignancies. Protocol 1 increased sensitivity and coverage of HaCaT proteome, it permitted us to reveal the provided new insights into the skin impact of SDS (widely used and rather non-toxic component of household chemicals, cosmetic products, shampoos and so on). Our results suggest the possible role of SDS in skin cancerogenesis.