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Qproteome Cell Compartment Kit
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咨询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.
Performance
See figures
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.
See figures
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.
Localization of proteins in cells under different growth conditions.
Micrographs (upper panels) of HeLa cells [A] under normal growth conditions and [B] after 4 hour incubation with staurosporine, an inducer of apoptosis. Lower panels show western blots prepared after cell lysates were processed using the Qproteome Cell Compartment Kit. The blots were probed with anti-cytochrome c antibody. It is known that during apoptosis cytochrome c is translocated from the intermembrane space of mitochondria to the cytosol, a process that is reflected in the detection of cytochrome c in the cytosolic fraction of apoptotic cells.
Specifications
| Features | Specifications |
|---|---|
| Applications | SDS-PAGE, mass spectrometry |
| Fractions isolated | Four fractions |
| Start material | Cell lysate |
| Sample size | ~5 x 10e6 cells |
| Binding capacity/yield | Varies |
| Species | Eukaryotes |