Bio-Rad ReadyPrep Protein Extraction Kit (Soluble/Insoluble) - Manual (Page 3)

Section 1
Introduction
The ReadyPrep protein extraction kit (Soluble/Insoluble) is
designed as a simple, rapid and reproducible method to
separate total cellular protein into two fractions, a soluble
fraction containing hydrophilic proteins and an insoluble
fraction containing hydrophobic proteins. This simple
fractionation strategy, which is based on differential protein
solubility, results in less complex samples and leads to
improved identification of low abundance proteins and a
better overall view of the proteome.
The short extraction protocol can be applied to a wide
variety of biological samples, from animal cells and tissues
to yeast, bacteria and plant tissues. The cells or tissue are
first suspended in Lysis Buffer and disrupted using an
ultrasonic probe. After centrifugation, the supernatant
containing the soluble hydrophilic proteins is separated
from the pellet containing the hydrophobic/insoluble
proteins. A second extraction of the pellet is performed
using additional lysis buffer to remove additional soluble
proteins and ensure complete cell lysis. The proteins in the
pellet are finally solubilized using 2-D rehydration/sample
buffer 1.
1
4110158A.qxd 6/16/2003 11:36 AM Page 4