Protein Array Patterning by Diffusive Gel Stamping

Proteins and small molecules are the effectors of physiological action in biological systems and comprehensive methods are needed to analyze their modifications, expression levels and interactions. Systems-scale characterization of the proteome requires thousands of components in high-complexity sam...

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Bibliographic Details
Main Authors: Anwar, Mekhail (Contributor), Gupta, Piyush (Contributor), Palaniapan, Raja (Contributor), Matsudaira, Paul T. (Contributor)
Other Authors: Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Biology (Contributor), Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science (Contributor), Whitehead Institute for Biomedical Research (Contributor)
Format: Article
Language:English
Published: Public Library of Science, 2013-01-23T15:08:19Z.
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Online Access:Get fulltext
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042 |a dc 
100 1 0 |a Anwar, Mekhail  |e author 
100 1 0 |a Massachusetts Institute of Technology. Department of Biological Engineering  |e contributor 
100 1 0 |a Massachusetts Institute of Technology. Department of Biology  |e contributor 
100 1 0 |a Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science  |e contributor 
100 1 0 |a Whitehead Institute for Biomedical Research  |e contributor 
100 1 0 |a Matsudaira, Paul T.  |e contributor 
100 1 0 |a Anwar, Mekhail  |e contributor 
100 1 0 |a Gupta, Piyush  |e contributor 
100 1 0 |a Palaniapan, Raja  |e contributor 
700 1 0 |a Gupta, Piyush  |e author 
700 1 0 |a Palaniapan, Raja  |e author 
700 1 0 |a Matsudaira, Paul T.  |e author 
245 0 0 |a Protein Array Patterning by Diffusive Gel Stamping 
260 |b Public Library of Science,   |c 2013-01-23T15:08:19Z. 
856 |z Get fulltext  |u http://hdl.handle.net/1721.1/76339 
520 |a Proteins and small molecules are the effectors of physiological action in biological systems and comprehensive methods are needed to analyze their modifications, expression levels and interactions. Systems-scale characterization of the proteome requires thousands of components in high-complexity samples to be isolated and simultaneously probed. While protein microarrays offer a promising approach to probe systems-scale changes in a high-throughput format, they are limited by the need to individually synthesize tens of thousands of proteins. We present an alternative technique, which we call diffusive gel (DiG) stamping, for patterning a microarray using a cellular lysate enabling rapid visualization of dynamic changes in the proteome as well protein interactions. A major advantage of the method described is that it requires no specialized equipment or in-vitro protein synthesis, making it widely accessible to researchers. The method can be integrated with mass spectrometry, allowing for the discovery of novel protein interactions. Here, we describe and characterize the sensitivity and physical features of DiG-Stamping. We demonstrate the biologic utility of DiG-Stamping by (1) identifying the binding partners of a target protein within a cellular lysate and by (2) visualizing the dynamics of proteins with multiple post-translational modifications. 
520 |a Whitaker Foundation 
546 |a en_US 
655 7 |a Article 
773 |t PLoS One