| Description: | The kit is designed for rapid fractionation of fresh/frozen soft plant tissues, especially leaves. Unlike other commercial kit and Lab protocols, of which large amount of starting material (a few grams) and lengthy processing time are required, this kit only uses 100-200 mg starting material. By using a specialized filter cartridge coupled with our proprietary buffer system, four subcellular fractions (cytosol, nuclei, chloroplasts and organelles) can be obtained in about 1 hour. The subcellular fractions can be used in numerous applications that include but not limited to protein trafficking, nuclear protein extraction and nucleic acid purification/sequencing. The highly enriched nuclei can be used for proteomics, flow cytometry analysis, DNA sequencing, and protein-protein/protein-DNA interaction studies. The subcellular fractionation protocol includes grinding tissue to release plant cells followed by disruption of the cell membrane by rapidly passing through the filter. Different subcellular fraction is obtained by differential centrifugation using only a table top centrifuge. This kit has been tested for use with many species such as Arabidopsis, Spinach, Peas, and Rape. |
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