An improved high throughput protein-protein interaction assay for nuclear hormone receptors.

Michael L. Goodson, Behnom Farboud, Martin L. Privalsky

Research output: Contribution to journalArticle

14 Citations (Scopus)

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 languageEnglish (US)
JournalNuclear receptor signaling
Volume5
StatePublished - 2007

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Cytoplasmic and Nuclear Receptors
Glutathione Transferase
Proteins
Centrifugation
Suspensions

ASJC Scopus subject areas

  • Medicine(all)

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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 journalArticle

Goodson, Michael L. ; Farboud, Behnom ; Privalsky, Martin L. / An improved high throughput protein-protein interaction assay for nuclear hormone receptors. In: Nuclear receptor signaling. 2007 ; Vol. 5.
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