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Life Science Research
Click to Enlarge 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.
ProteoExtract® Subcellular Proteome Extraction Kit (S-PEK) is designed for fast and reproducible extraction of subcellular proteomes from mammalian tissue and adherent and suspension-grown cells. S-PEK takes advantage of the different solubilities of certain subcellular compartments in the four selected reagents.
The ProteoExtract® S-PEK™ Antibody Control Kit is a set of four monoclonal antibodies that recognize proteins specific for each of the four sub-cellular fractions obtained with the ProteoExtract® Subcellular Proteome Extraction Kit (S-PEK). The ProteoExtract® Complete Mammalian Proteome Extraction Kit (C-PEK) designed to extract virtually all proteins from both mammalian tissues and culture cells in a simple two step process within a microcentrifuge tube to minimize protein loss. The ProteoExtract® Cytosol/Mitochondria Fractionation Kit is designed for extraction of cellular proteomes from mammalian cells. The Merck ProteoExtract® Cytoskeleton Enrichment and Isolation Kit and the ProteoExtract® Native Cytoskeleton Enrichment and Staining Kit provides cytoskeleton purification detergent buffers, sufficient for extraction from ten 100 mm culture dishes, that retain focal adhesion and actin-associated proteins while removing soluble cytoplasmic and nuclear proteins from the cell.