Abstract
The analysis of EpiDerm TM cultures treated with the known skin irritant sodium lauryl sulphate (SLS) was performed using 2D-gel electrophoresis in order to understand the mechanism of action and thereby identify novel markers of skin irritation. A range of both broad and narrow pH gradient first-dimension gels were run (pH 4–7, 6–11, 4–5, 5–6 and 6–9) consistently followed by 12% SDS-PAGE in the second-dimension. Following treatment of EpiDerm TM with SLS, 67 proteins of interest were identified, of which 8 were selected as interesting: calmodulin-like skin protein, involucrin, epithelial cell marker protein, HS1, peroxiredoxin 1, serine protease inhibitor, KIAA0117 and ribosomal protein L17. Involucrin was confirmed as being up-regulated by both ELISA and Western blotting. The use of proteomics has identified a number of proteins which could be used as general markers for skin irritation and which may in particular be of value for the development of in vitro predictive models.
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