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Efficient Cellular Fractionation for RNA ,,, and Protein Isolation

Efficient Cellular Fractionation for RNA and Protein Isolation


Ambion's Protein and RNA Isolation System, PARIS, uses a rapid and simple procedure to isolate both total RNA and native protein from the same experimental sample. Cultured cells and tissues that are either fresh, frozen, or treated with RNA stabilization reagents such as RNAlater and RNAlater-ICE, are all compatible with the procedure. This kit provides a unique tool for researchers who need both RNA and protein for various downstream analysis, reducing time, cost, and variability between independent experimental samples. The PARIS Kit also permits separation of nuclear and cytoplasmic fractions from fresh cultured cells prior to RNA and/or protein isolation. Here, we tested the procedure with various mammalian cell types and show that both RNA and native protein are efficiently recovered from nuclear and cytoplasmic fractions with virtually no cross contamination between the two cellular compartments.


Fast and Simple Cellular Fractionation
To obtain total cellular protein and RNA using the PARIS Kit, whole cells are first homogenized in ice-cold Cell Disruption Buffer to prepare a total cell lysate. Since the homogenization is performed quickly on ice and in the presence of detergent, both protein and high quality RNA can be isolated from this lysate. For RNA isolation, a part of the tota
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