Simultaneous extraction of nucleic acids and proteins from tissue specimens by ultracentrifugation: A protocol using the high-salt protein fraction for quantitative proteome analysis.

Abstract:

:Comprehensive molecular profiling of human tumor tissue specimens at the DNA, mRNA and protein level is often obstructed by a limited amount of available material. Homogenization of frozen tissue samples in guanidine isothiocyanate followed by ultracentrifugation over cesium chloride allows the simultaneous extraction of high-molecular weight DNA and RNA. Here, we present a protocol for quantitative proteome analysis using the high-salt protein fraction obtained as supernatant after ultracentrifugation for nucleic acid extraction. We applied this method to extracts from primary human brain tumors and demonstrate its successful application for protein expression profiling in these tumors using 2-D DIGE, MS and Western blotting.

journal_name

Proteomics

journal_title

Proteomics

authors

Grzendowski M,Riemenschneider MJ,Hawranke E,Stefanski A,Meyer HE,Reifenberger G,Stühler K

doi

10.1002/pmic.200800902

subject

Has Abstract

pub_date

2009-11-01 00:00:00

pages

4985-90

issue

21

eissn

1615-9853

issn

1615-9861

journal_volume

9

pub_type

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