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Related Concept Videos

DNA Microarrays02:34

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...

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A HaloTag-based small molecule microarray screening methodology with increased sensitivity and multiplex

Devin J Noblin1, Charlotte M Page, Hyun Seop Tae

  • 1Department of Molecular Cellular and Developmental Biology, Yale University, 219 Prospect Street, New Haven, CT 06511, USA.

ACS Chemical Biology
|September 28, 2012
PubMed
Summary

This study enhances small molecule microarrays (SMMs) using HaloTag technology for increased sensitivity and multiplex screening. This novel method enables precise identification of specific protein isoform ligands.

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Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology
11:16

Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology

Published on: July 12, 2014

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Chemical Biology

Background:

  • Small Molecule Microarrays (SMMs) are valuable for screening small molecule-protein interactions.
  • Current SMM methods have limitations in sensitivity and multiplexing capabilities.

Purpose of the Study:

  • To enhance SMM methodology for increased assay sensitivity.
  • To develop a multiplex screening approach for SMMs.
  • To enable the identification of isoform-specific ligands.

Main Methods:

  • Fusion of target proteins to HaloTag for covalent prelabeling with fluorophores.
  • Application of HaloTag-based SMMs to screen FKBP12-ligand interactions.
  • Utilizing different fluorophores for multiplex screening of protein isoforms (e.g., wild type FKBP12 and FKBP12 F36V mutant).
  • Screening a 20,000 compound library against PTP1B isoforms.

Main Results:

  • HaloTag-based SMMs significantly increased assay sensitivity for FKBP12-ligand interactions.
  • Multiplex screening successfully differentiated ligands for wild type FKBP12 and the F36V mutant.
  • The technique identified ligands selective for a specific PTP1B isoform from a large compound deck.

Conclusions:

  • HaloTag fusion enhances SMM sensitivity and enables robust multiplex screening.
  • This modified SMM platform facilitates the discovery of isoform-specific small molecule ligands.
  • The methodology holds promise for drug discovery and chemical biology research.