ProteoGrade™ Cell Compartment Kit

The ProteoGrade™ Cell Compartment Kit offers a comprehensive solution for subcellular fractionation, enabling precise localization of proteins within eukaryotic cells and tissues. By sequentially applying specialized extraction buffers, proteins from distinct cellular compartments such as cytosol, membranes, nucleus, and cytoskeleton can be selectively isolated, facilitating a deeper understanding of cellular organization and function.

This kit simplifies the process of subcellular fractionation, allowing for efficient extraction and isolation of proteins associated with specific cellular compartments. Its performance is demonstrated by the clear and specific separation of marker proteins, both in isolated cells and tissue samples, as illustrated in the accompanying figures.

The principle behind the ProteoGrade™ Cell Compartment Kit lies in the sequential addition of four extraction buffers to a cell pellet or tissue sample. Each buffer targets proteins residing in different cellular compartments, enabling their selective extraction and subsequent analysis.

The procedure is straightforward: following the sequential addition of extraction buffers to the cell pellet, fractions are isolated through centrifugation. For tissue samples, an alternative protocol involving simultaneous homogenization and disruption using the TissueRuptor is employed before processing akin to cell pellets.

Subcellular fractionation facilitated by this kit serves various applications, including the enrichment of low-abundance species, defining the subcellular localization of enzymes and regulatory proteins, and monitoring compartmental redistribution of biomolecules under different physiological conditions.

Supporting data showcases the effectiveness of the kit in localizing proteins under diverse growth conditions, offering valuable insights into cellular processes such as apoptosis-induced redistribution of cytochrome c. In terms of specifications, the kit is compatible with SDS-PAGE and mass spectrometry, capable of isolating four distinct fractions from a starting material of cell lysate, with a typical sample size of approximately 5 x 10^6 cells. The binding capacity and yield vary depending on the specific proteins and cellular compartments of interest, with applications tailored for eukaryotic species.