The Thermo Scientific Subcellular Protein Isolation Kit for cultured cells isolates and enriches proteins from 5 different cellular compartments. Collect protein.

Subcellular Protein Isolation Kit for Cultured Cells Features:

•Simple — Extract functionality from mammalian cultured cells in less than 3 hours Cytoplasmic, membrane, soluble nuclear, chromatin-binding, and cytoskeletal protein components
• Convenience — perform simple procedures without gradient ultracentrifugation
• Complete Set b> — Obtain cytoplasmic, membrane, soluble nuclear, chromatin-binding, and cytoskeletal protein fractions using a single kit
• Cultured Cells — Developed and optimized for use with cultured mammalian cells ;For tissue samples, use Subcellular Isolation Kit for Tissue
• Compatible—Use extracts for downstream applications such as protein detection, Western blotting, protein detection, electrophoretic mobility shift assays, reporter gene and enzyme activity assays.

Applications of subcellular fractions:
• Extraction and enrichment of proteins from different cellular compartments
• Isolation of cytoplasm, nucleus, membrane and Cytoskeletal Proteins
• Analyze proteins in the cytoplasm, nucleus, membrane, and cytoskeleton
• Study protein translocation

Subcellular protein isolation enables assessment of protein localization and proteins from specific cellular compartments Enrichment. Thermo Scientific Subcellular Protein Isolation Kits contain reagents for step-by-step lysis of cells into functional cytoplasmic, membrane, soluble nuclear, chromatin-bound, and cytoskeletal protein fractions in less than 3 hours. Extracts obtained from each subcellular compartment typically have less than 15% contamination between fractions, a purity sufficient for most experiments investigating protein localization and redistribution.

Subcellular Protein Isolation Kit for Cultured Cells contains 4 extraction buffers, a stable nuclease, and Thermo Scientific Halt Protease Inhibitor Cocktail. Each kit contains sufficient reagents to isolate 50 20 µL cell pellets (equivalent to approximately 2 g of cell pellets). (For tissue samples, use our Subcellular Protein Isolation Kit for Tissue.)

The first kit reagent added to the cell pellet causes selective membrane permeabilization, releasing soluble cytoplasmic contents. The second reagent dissolves plasma, mitochondria, and ER-Golgi membranes, but not nuclear membranes. After recovering intact nuclei by centrifugation, the third reagent yields a soluble nuclear extract. An additional nuclear extraction using micrococcal nuclease is performed to release chromatin-linked nuclear proteins. The recovered insoluble pellets are then extracted using a final reagent to isolate cytoskeletal proteins.

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