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...
| Main Authors: | , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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Public Library of Science
2013
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| Online Access: | http://hdl.handle.net/1721.1/76339 https://orcid.org/0000-0002-9703-1780 |
| _version_ | 1826202655307857920 |
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| author | Anwar, Mekhail Gupta, Piyush Palaniapan, Raja Matsudaira, Paul T. |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Anwar, Mekhail Gupta, Piyush Palaniapan, Raja Matsudaira, Paul T. |
| author_sort | Anwar, Mekhail |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T12:12:40Z |
| format | Article |
| id | mit-1721.1/76339 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T12:12:40Z |
| publishDate | 2013 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | mit-1721.1/763392022-09-28T00:43:32Z Protein Array Patterning by Diffusive Gel Stamping Anwar, Mekhail Gupta, Piyush Palaniapan, Raja Matsudaira, Paul T. Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Whitehead Institute for Biomedical Research Matsudaira, Paul T. Anwar, Mekhail Gupta, Piyush Palaniapan, Raja 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. Whitaker Foundation 2013-01-23T15:08:19Z 2013-01-23T15:08:19Z 2012-10 2012-04 Article http://purl.org/eprint/type/JournalArticle 1932-6203 http://hdl.handle.net/1721.1/76339 Anwar, Mekhail et al. “Protein Array Patterning by Diffusive Gel Stamping.” Ed. Rizwan Hasan Khan. PLoS ONE 7.10 (2012): e46382. Web. https://orcid.org/0000-0002-9703-1780 en_US http://dx.doi.org/10.1371/journal.pone.0046382 PLoS One Creative Commons Attribution http://creativecommons.org/licenses/by/2.5/ application/pdf Public Library of Science PLoS |
| spellingShingle | Anwar, Mekhail Gupta, Piyush Palaniapan, Raja Matsudaira, Paul T. Protein Array Patterning by Diffusive Gel Stamping |
| title | Protein Array Patterning by Diffusive Gel Stamping |
| title_full | Protein Array Patterning by Diffusive Gel Stamping |
| title_fullStr | Protein Array Patterning by Diffusive Gel Stamping |
| title_full_unstemmed | Protein Array Patterning by Diffusive Gel Stamping |
| title_short | Protein Array Patterning by Diffusive Gel Stamping |
| title_sort | protein array patterning by diffusive gel stamping |
| url | http://hdl.handle.net/1721.1/76339 https://orcid.org/0000-0002-9703-1780 |
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