Reconstituted Epidermis timecourse of barrier formation

Data from PeptideAtlas, [[http://www.peptideatlas.org/PASS/PASS00363 PASS00363]]. Experiment: P2-D3, file: P2-D3-F6.mzml. From PeptideAtlas: {{i}} De-cellularized dermis was seeded with neonatal keratinocytes and cultured submerged in media for two days. At this point the cultures were raised to the air/water interface to stimulate differentiation. At the indicated time points, epidermis was removed with forceps. Epidermis was extracted in 50% Trifluoroethanol (TFE), 0.1% SDS, 100 mM Ammonium Bicarbonate (AMBIC). Samples were heated at 60 deg C for 30 minutes, then sonicated at 75 W for 2 minutes process time using a 50% duty cycle. Extracts were cleared by centrifugation. separation: Samples from each timepoint were randomly assigned to 4 pools. Pools of 250 ug peptides were separated with Offgel Electrophoresis, pH 3-10 into 12 fractions. {{/i}}