Abstract
Various assays have been employed to study the nuclear receptor/cofactor interactions. Coimmunoprecipitation protocols, both yeast and mammalian two-hybrid systems, and electrophoretic mobility shift/supershift assays are all commonly used. One of the most useful assays for studying direct protein-protein interactions is the glutathione-S-transferase "pulldown" assay. We have developed a high-throughput version of this assay that utilizes a filter microplate to allow parallel processing of many samples, significantly reducing the time and reagents required for the assay and increasing the sensitivity of the assay for weaker protein-protein interactions.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 157-169 |
| Number of pages | 13 |
| Journal | Methods in molecular biology (Clifton, N.J.) |
| Volume | 505 |
| DOIs | |
| State | Published - 2009 |
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ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
High throughput analysis of nuclear receptor-cofactor interactions. / Goodson, Michael L.; Farboud, Behnom; Privalsky, Martin L.
In: Methods in molecular biology (Clifton, N.J.), Vol. 505, 2009, p. 157-169.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - High throughput analysis of nuclear receptor-cofactor interactions.
AU - Goodson, Michael L.
AU - Farboud, Behnom
AU - Privalsky, Martin L.
PY - 2009
Y1 - 2009
N2 - Various assays have been employed to study the nuclear receptor/cofactor interactions. Coimmunoprecipitation protocols, both yeast and mammalian two-hybrid systems, and electrophoretic mobility shift/supershift assays are all commonly used. One of the most useful assays for studying direct protein-protein interactions is the glutathione-S-transferase "pulldown" assay. We have developed a high-throughput version of this assay that utilizes a filter microplate to allow parallel processing of many samples, significantly reducing the time and reagents required for the assay and increasing the sensitivity of the assay for weaker protein-protein interactions.
AB - Various assays have been employed to study the nuclear receptor/cofactor interactions. Coimmunoprecipitation protocols, both yeast and mammalian two-hybrid systems, and electrophoretic mobility shift/supershift assays are all commonly used. One of the most useful assays for studying direct protein-protein interactions is the glutathione-S-transferase "pulldown" assay. We have developed a high-throughput version of this assay that utilizes a filter microplate to allow parallel processing of many samples, significantly reducing the time and reagents required for the assay and increasing the sensitivity of the assay for weaker protein-protein interactions.
UR - http://www.scopus.com/inward/record.url?scp=60549098518&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=60549098518&partnerID=8YFLogxK
U2 - 10.1007/978-1-60327-575-0_9
DO - 10.1007/978-1-60327-575-0_9
M3 - Article
VL - 505
SP - 157
EP - 169
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
SN - 1064-3745
ER -