Bio-Rad ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear) User Manual
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Section 1
Introduction
The ReadyPrep protein extraction kit (cytoplasmic/nuclear)
is designed to quickly prepare highly enriched fractions of
cytoplasmic and nuclear proteins from eukaryotic samples,
such as cultured cells and tissues. This kit is part of a
group of related kits from Bio-Rad developed to reduce
sample complexity in order to improve the chances of
identifying low abundant proteins and to simplify proteomic
studies. Although this kit is directed towards the
preparation of nuclear and cytoplasmic protein samples for
2-D gel electrophoresis, the fractions are suitable for many
applications such as SDS-PAGE and western blotting.
The ReadyPrep protein extraction kit (cytoplasmic/nuclear)
utilizes a specially fomulated buffer and differential
centrifugation to isolate intact nuclei (Dignam et al. 1983,
Zerivitz and Akusjarvi 1989). The kit also contains a
strongly chaotropic extraction buffer to efficiently solubilize
nuclear proteins for immediate use in 2-D gel
electrophoretic applications. Analysis of the cytoplasmic
fraction requires that the protein first be processed through
the ReadyPrep 2-D cleanup kit, which is provided. This kit
removes components from the extraction buffer that would
interfere with the isoelctric focusing step.
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