RNA and protein clean-up from the same specimen. Comparison between the Qiagen and Ambion protocols

Authors: Prestvik, W. S.¹; Hjertø, A. L.¹; Steigedal, T. S.²; Thommesen, L.³

Source: Scandinavian Journal of Clinical and Laboratory Investigation, Volume 67, Number 8, 2007 , pp. 885-891(7)

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Abstract:

Study of gene and protein expression patterns is considered desirable for gaining new insight into cellular molecular mechanisms. The purpose of the present study was to compare the RNeasy (Qiagen) and PARIS (Ambion) protocols for protein and RNA clean-up. We found that the RNeasy protocol resulted in a significantly higher yield of both RNA and protein compared to the PARIS protocol. Western blot analysis of proteins isolated with these two methods, and with use of three different antibodies (PKB/Akt, c-fos and CREM), indicated that these methods were equal in regard to protein patterns, despite the protein precipitation step that is included in the RNeasy protocol. In some cases, Western blot performed on unprecipitated flow-through (Qiagen) may be satisfactory for Western blot analysis, but the amount of protein is probably decisive for detection.

Keywords: AR42J; MC3T3-E1; PARIS; protein isolation; RNA isolation; RNeasy

Document Type: Research article

DOI: http://dx.doi.org/10.1080/00365510701432491

Affiliations: 1: Department of Food Science and Medical Technology, Sør-Trøndelag University College, Trondheim, Norway 2: Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway 3: Department of Food Science and Medical Technology, Sør-Trøndelag University College, Trondheim, Norway,Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway

Publication date: 2007-01-01

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RNA and protein clean-up from the same specimen. Comparison between the Qiagen and Ambion protocols

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