Qiagen, 37502, Qproteome Cell Compartment Kit

For up to 10 subcellular fractionations: Extraction Buffer, Protease Inhibitor Solution, Benzonase®.

Product Details

Eukaryotic cells are complex, well-ordered, and highly structured systems. The Cell Compartment Kit is designed for fast and easy subcellular fractionation of intact eukaryotic cells and tissue. By sequential addition of different extraction buffers to a cell pellet, proteins in the different cellular compartments can be selectively isolated.

Principle

The kit contains 4 Extraction Buffers which enable the sequential isolation of proteins associated with the cytosol, membranes, nucleus, and cytoskeleton from cell lysates or tissues.

Procedure

The 4 Extraction Buffers are added sequentially to a cell pellet and the respective fractions are isolated by centrifugation (see flowchart ).

In an alternative protocol for tissues, samples are simultaneously homogenized and disrupted using the TissueRuptor before being processed as for the cell pellet.

Applications

Subcellular fractionation of proteins can be used for the enrichment of low-abundance species; to define the subcellular localization of enzymes, regulatory, and structural proteins; and for monitoring of compartmental redistribution of biomolecules under basal and stimulated conditions.