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Abcam's Cell Fractionation Kit - High Throughput (HT), provides a method and reagents for a simple and rapid preparation of cytosolic, mitochondrial and nuclear fractions from adherent cells.
The Cell Fractionation Kit is designed to produce highly enriched fractions that allow for the monitoring of proteins of interest through various cellular compartments. It is not designed to produce purified cellular fractions.
The kit is especially suitable for, but not limited to, high throughput fractionation of adherent cells in a 96-well plate format. The entire fractionation of 96 samples into the cytosolic, mitochondrial and nuclear fraction is achieved in one hour. The kit is based on sequential detergent extraction of cytosolic, mitochondrial and nuclear proteins without the need for mechanical disruption of cells, and thus fractionates adherent cells into cytosol-containing, mitochondria-containing and nuclei-containing fractions. These fractions are referred throughout the protocol as cytosolic, mitochondrial and nuclear fractions. The kit prepares sufficient sample material for subsequent Western blot analyses, dot blot analyses or for analyses by microplate ELISA assays.
ab109718 is designed to allow the measurement of any proteins which are differentially represented in the cytosol, mitochondria and nuclei, and is particularly applicable to studies of proteins that translocate between these three cellular compartments.
A second version of the kit is also available, ab109719 Cell Fractionation Kit - Standard. This version is designed for fractionating cells grown in batches and is ideal for experiments requiring larger sample amounts.
Storage: Store Buffer A, Detergent HT II, Detergent HT III and 5X SDS Sample Buffer at -20°C. Store Detergent HT I at -80°C. Store the collection plates and seal films at room temperature.
|Components||1 x 96 tests|
|5X SDS Sample Buffer||1 x 1.5ml|
|96-well collection plate with seal film||3 units|
|Buffer A||1 x 34ml|
|Detergent HT I||1 x 7.5µl|
|Detergent HT II||1 x 35µl|
|Detergent HT III||1 x 700µl|
|Plate Seals||3 units|
Our Abpromise guarantee covers the use of ab109718 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent dilution.|