Thermo Scientific™ Subcellular Protein Fractionation Kit for Cultured Cells

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 for Cultured Cells 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:

• Concise – extract functional cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal protein fractions in less than three 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

• Cultured Cells – developed and optimized for use with cultured mammalian cells; for tissue samples use the Subcellular Fractionation Kit for Tissues, Part No. 87790

• Compatible – use with a variety of downstream applications including Western blotting, protein assays, electrophoretic mobility shift assays and reporter-gene and enzyme-activity assays

• Small-scale – kit provides sufficient reagents to fractionate 50 cell pellets of 20μL each, equivalent to a total of 2g of cell paste or tissue

Includes:

Four extraction buffers, micrococcal nuclease enzyme, calcium chloride solution and protease inhibitor cocktail

Recommended for:

Extraction and enrichment of proteins from different cellular compartments; Separation and profiling of protein from the cytoplasm, nucleus, membrane and cytoskeleton from the same cell sample; Studying protein translocation