( l cell lysate is i...)Efficient Cellular Fractionation for RNA ,,, and Protein Isolation

l cell lysate is immediately mixed with an equal volume of Lysis/Binding Solution. This solution contains a high concentration of guanidinium thiocyanate, a strong chaotropic denaturant that rapidly inactivates cellular ribonucleases. Total RNA is then purified from the mixture using a RNA binding glass fiber filter. After three rapid washing steps, high quality, concentrated RNA is eluted.

The PARIS Kit also contains a solution optimized for rapid nuclear/cytoplasmic fractionation of fresh cultured cells. The fractionation method is based on differential lysis of plasma and nuclear membranes by nonionic detergents. After selective lysis of the plasma membrane in Cell Fractionation Buffer, intact nuclei are collected by a quick centrifugation step and resuspended in Cell Disruption Buffer. The supernatant contains all of the cytoplasmic components. Following the same procedure described above, both protein and high quality RNA can then be isolated from each cellular fraction. The entire procedure for the simultaneous isolation of RNA and native protein from total or nuclear and cytoplasmic fractions can be completed in as little as 30 minutes.

Efficient Cellular Fractionation with Various Cell Types
To examine the efficiency of the PARIS procedure, nuclear and cytoplasmic protein and RNA were isolated from 1x10⁶ HEK 293 cells in triplicate and compared to total protein and RNA isolated from the same number of cells (Fi
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