Bio-Rad ReadyPrep™ Protein Extraction Kit (Membrane II) User Manual

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Section 1
Introduction

The ReadyPrep protein extraction kit (membrane II) is
designed as a simple, rapid and reproducible method to
isolate cellular membranes and then to extract the
associated integral membrane proteins into a solution
compatible with two-dimensional polyacrylamide gel
electrophoresis (2-D PAGE). The ReadyPrep protein extrac-
tion kit (membrane II) joins two existing ReadyPrep kits for
isolation of membrane proteins; it is optimized to effectively
isolate more complex proteins that contain more than two
transmembrane domains, or are resistant to extraction by
the ReadyPrep membrane I kit. The signal kit is specific for
isolation of proteins involved in intracellular membrane
trafficking and signaling pathways. All three of these mem-
brane kits reduce sample complexity using cellular location
as a way to fractionate. The procedure for the ReadyPrep
protein extraction kit (membrane II) is based on the original
work of Fujiki et al.

1

for the isolation of cellular membrane

fractions from rat liver after carbonate extraction and the
later adaption of the technique by Molloy et al.

2

for

Proteomic analysis of E. coli outer membrane proteins.

The kit contains all the reagents necessary for membrane
isolation and membrane-protein extraction and the protocol

1

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