Subcellular Protein Fractionation Kit Summary
| Description |
The Cell Fractionation kit is designed to provide reproducible extraction of four subcellular protein fractions (Cytosol, nucleus, membrane/particulate, and cytoskeletal fractions) from a single mammalian sample. The method is fast and simple, needing only 2 hours and no ultracentrifugation involved. All four protein fractions obtained are suitable for many downstream applications such as 1-D or 2-D gel, enzyme activity assays, gel shift assay, and Western blotting.
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| Kit Type |
Kit
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Applications/Dilutions
| Application Notes |
The Cell Fractionation Kit can be used in 1-D or 2-D gel, enzyme activity assays, gel shift assay, and Western blotting. In Western analysis, if a membrane protein appeares mainly in the nuclear fraction, the membrane extraction step may not be sufficient. First, make sure to add Membrane Extraction Buffer B into the extraction, which contain NP-40 detergent. If it still shows the same result, increase incubation time from 1 min to 5 min after adding MEB-B. NP-40 should extract membrane from most of cells.
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Packaging, Storage & Formulations
| Storage |
Store at -20C. Avoid freeze-thaw cycles.
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Kit Components
| Components |
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Notes
The percentage of proteins in the different fractions are cell-dependent, usually in the range of ~20-30% cytosol, ~25-35% membrane (organelles), ~15-25% Nuclei, ~15-25% insoluble