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
Proteomicsjournal_title
Proteomicsauthors
Grzendowski M,Riemenschneider MJ,Hawranke E,Stefanski A,Meyer HE,Reifenberger G,Stühler Kdoi
10.1002/pmic.200800902subject
Has Abstractpub_date
2009-11-01 00:00:00pages
4985-90issue
21eissn
1615-9853issn
1615-9861journal_volume
9pub_type
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