single bead affinity detection (sinbad) for the analysis of protein-protein interactions单珠亲和检测(辛巴达)分析蛋白质相互作用.pdf

Single Bead Affinity Detection (SINBAD) for the Analysis of Protein-Protein Interactions Roberta Schulte, Jessica Talamas, Christine Doucet, Martin W. Hetzer* Molecular and Cell Biology Laboratory, Salk Institute for Biological Studies, La Jolla, California, United States of America Abstract We present a miniaturized pull-down method for the detection of protein-protein interactions using standard affinity chromatography reagents. Binding events between different proteins, which are color-coded with quantum dots (QDs), are visualized on single affinity chromatography beads by fluorescence microscopy. The use of QDs for single molecule detection allows the simultaneous analysis of multiple protein-protein binding events and reduces the amount of time and material needed to perform a pull-down experiment. Citation: Schulte R, Talamas J, Doucet C, Hetzer MW (2008) Single Bead Affinity Detection (SINBAD) for the Analysis of Protein-Protein Interactions. PLoS ONE 3(4): e2061. doi:10.1371/journal.pone.0002061 Editor: Edathara Abraham, University of Arkansas, United States of America Received March 5, 2008; Accepted March 12, 2008; Published April 30, 2008 Copyright: 2008 Hetzer et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: Innovation Grant, Salk Institute. This work was supported by the Pew Scholar program and NIH R01 GM073994. Competing Interests: The authors have declared that no competing interests exist. * E-mail: hetzer@ Introduction the TAP-tagged protein. The distribution of fluorescence intensity