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Subcellular Protein Fractionation Kit
Segregate and enrich proteins from five cellular compartments.
The Thermo Scientific Subcellular Protein Fractionation Kit enables segregation and enrichment of proteins from five different cellular compartments.
The kit enables the stepwise separation and extraction of cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal proteins from mammalian cultured cells. Extracts obtained with the Subcellular Protein Fractionation Kit are compatible with a variety of downstream applications including Western blotting, protein assays, electrophoretic mobility shift assays and reporter-gene and enzyme-activity assays.
Fractionation of subcellular proteins enables protein localization assessment and protein enrichment from specific cellular compartments. The Thermo Scientific Subcellular Protein Fractionation Kit includes a combination of reagents for stepwise lysis of cells into functional cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal protein fractions in less than 3 hours. Extracts from each subcellular compartment generally have less than 15% contamination between fractions, which is sufficient purity for most experiments studying protein localization and redistribution.
Highlights of the Subcellular Fractionation Kits:
Concise – extract functional cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal protein fractions in less than 3 hours
Convenient – perform a simple procedure without using ultracentrifugation over gradients
Complete – obtain cytoplasmic, membrane, soluble nuclear, chromatin-bound and cytoskeletal protein fractions from a single kit
Compatible – use extracts for downstream applications such as protein assays, Western blotting, gel-shift assays and enzyme activity assays
Applications for subcellular fractionation:
Extract and enrich proteins from different cellular compartments
Isolate cytoplasmic, nuclear, membrane and cytoskeletal proteins from the same cells
Profile cells for proteins from the cytoplasm, nucleus, membrane and cytoskeleton
Study protein translocation
Product Details:
The Subcellular Protein Fractionation Kit contains four extraction buffers, a stabilized nuclease and Thermo Scientific Halt Protease Inhibitor Cocktail. Each kit has enough reagents to fractionate 50 cell pellets of 20µL each, equivalent to approximately 2g of cell paste. The first reagent added to a cell pellet causes selective membrane permeabilization, releasing soluble cytoplasmic contents. The second reagent dissolves plasma, mitochondria and ER-golgi membranes but does not solubilize the nuclear membranes. After recovering intact nuclei by centrifugation, a third reagent yields the soluble nuclear extract. An additional nuclear extraction with micrococcal nuclease is performed to release chromatin-bound nuclear proteins. The recovered insoluble pellet is then extracted with the final reagent to isolate cytoskeletal proteins.
Subcellular Protein Fractionation Kit
Formulation: Multi-component kit
Sufficient For: 50 samples each containing 2 million cells (20µL packed)
Kit Contents:
Cytoplasmic Extraction Buffer, 10mL
Membrane Extraction Buffer, 10mL
Nuclear Extraction Buffer, 10mL
Pellet Extraction Buffer, 5mL
Micrococcal Nuclease, 150µL
Calcium Chloride, 100mM, 250µL
Halt Protease Inhibitor Cocktail (100X), 350µL |