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Morphological changes of cells during subcellular extraction
A431 cells were incubated with Phallicidin, DAPI and MitoTracker in order to visualize actin filaments, nuclei and mitochondria, respectively. Stepwise extraction of cytosolic fraction (F1), organelle/membrane fraction (F2), nuclear fraction (F3) and cytoskeletal fraction (F4) monitored by fluorescence microscopy shows sequential degradation of cellular structure.
2DE-Gel with 250 µg Protein from HepG2 liver carcinoma cells. Cells were collected by centrifugation and sample preparation was performed. 200 µg proteins of the whole cell extract were separated by 2DE. First dimension was done by isoelectric focusing from pH 4 to pH 7 in an immobilized pH gradient gel by application of 50.000 Vh. The second dimension separation was by SDS-PAGE in a 12 % polyacrylamid gel. Proteins were visualized by silver staining.