Abstract
The Glutathione-S-Transferase (GST) "pulldown" assay has been used extensively to assay protein interactions in vitro. This methodology has been especially useful for investigating the interactions of nuclear hormone receptors with a wide variety of their interacting partners and coregulatory proteins. Unfortunately, the original GST-pulldown technique relies on multiple binding, washing and elution steps performed in individual microfuge tubes, and requires repeated centrifugation, aspiration, and suspension steps. This type of batch processing creates a significant liquid handling bottleneck, limiting the number of sample points that can be incorporated into one experiment and producing inherently less efficient washing and elution than would a flow-through methodology. In this manuscript, we describe the adaptation of this GST-pulldown assay to a 96-well filter plate format. The use of a multi-well filter plate makes it possible to assay more samples in significantly less time using less reagents and more efficient sample processing than does the traditional single tube assay.
| Original language | English (US) |
|---|---|
| Journal | Nuclear receptor signaling |
| Volume | 5 |
| State | Published - 2007 |
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ASJC Scopus subject areas
- Medicine(all)
Cite this
An improved high throughput protein-protein interaction assay for nuclear hormone receptors. / Goodson, Michael L.; Farboud, Behnom; Privalsky, Martin L.
In: Nuclear receptor signaling, Vol. 5, 2007.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - An improved high throughput protein-protein interaction assay for nuclear hormone receptors.
AU - Goodson, Michael L.
AU - Farboud, Behnom
AU - Privalsky, Martin L.
PY - 2007
Y1 - 2007
N2 - The Glutathione-S-Transferase (GST) "pulldown" assay has been used extensively to assay protein interactions in vitro. This methodology has been especially useful for investigating the interactions of nuclear hormone receptors with a wide variety of their interacting partners and coregulatory proteins. Unfortunately, the original GST-pulldown technique relies on multiple binding, washing and elution steps performed in individual microfuge tubes, and requires repeated centrifugation, aspiration, and suspension steps. This type of batch processing creates a significant liquid handling bottleneck, limiting the number of sample points that can be incorporated into one experiment and producing inherently less efficient washing and elution than would a flow-through methodology. In this manuscript, we describe the adaptation of this GST-pulldown assay to a 96-well filter plate format. The use of a multi-well filter plate makes it possible to assay more samples in significantly less time using less reagents and more efficient sample processing than does the traditional single tube assay.
AB - The Glutathione-S-Transferase (GST) "pulldown" assay has been used extensively to assay protein interactions in vitro. This methodology has been especially useful for investigating the interactions of nuclear hormone receptors with a wide variety of their interacting partners and coregulatory proteins. Unfortunately, the original GST-pulldown technique relies on multiple binding, washing and elution steps performed in individual microfuge tubes, and requires repeated centrifugation, aspiration, and suspension steps. This type of batch processing creates a significant liquid handling bottleneck, limiting the number of sample points that can be incorporated into one experiment and producing inherently less efficient washing and elution than would a flow-through methodology. In this manuscript, we describe the adaptation of this GST-pulldown assay to a 96-well filter plate format. The use of a multi-well filter plate makes it possible to assay more samples in significantly less time using less reagents and more efficient sample processing than does the traditional single tube assay.
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UR - http://www.scopus.com/inward/citedby.url?scp=34249654613&partnerID=8YFLogxK
M3 - Article
C2 - 17464356
AN - SCOPUS:34249654613
VL - 5
JO - Nuclear receptor signaling
JF - Nuclear receptor signaling
SN - 1550-7629
ER -