Open AccessProbing Small‐Molecule Microarrays with Tagged Proteins in Cell Lysates
Author(s) -
Pop Marius S.,
Wassaf Dina,
Koehler Angela N.
Publication year - 2014
Publication title -
current protocols in chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.503
H-Index - 14
ISSN - 2160-4762
DOI - 10.1002/9780470559277.ch140101
Subject(s) - protein array analysis , small molecule , protein microarray , context (archaeology) , computational biology , lysis , fluorescence , chemistry , high throughput screening , target protein , microarray , protein–protein interaction , microbiology and biotechnology , dna microarray , biochemistry , biology , gene expression , gene , paleontology , physics , quantum mechanics
The technique of small‐molecule microarray (SMM) screening is based on the ability of small molecules to bind to various soluble proteins. This type of interaction is easily detected by the presence of a fluorescence signal produced by labeled antibodies that specifically recognize a unique sequence (tag) present on the target protein. The fluorescent signal intensity values are determined based on signal‐to‐noise ratios (SNRs). SMM screening is a high‐throughput, unbiased method that can rapidly identify novel direct ligands for various protein targets. This binding‐based assay format is generally applicable to most proteins, but it is especially useful for protein targets that do not possess an enzymatic activity. SMMs enable screening a protein in a purified form or in the context of a cellular lysate, likely providing a more physiologically relevant screening environment. © 2014 by John Wiley & Sons, Inc.