Small-molecule reagents for cellular pull-down experiments.
Affinity purification of interacting proteins from cellular extracts is a powerful technique for identifying the cellular targets of small molecules. Affinity matrix-based small-molecule reagents are usually prepared by conjugating small molecules of interest to a solid support such as agarose. This protocol describes an efficient and robust method to immobilize small molecules containing a primary alcohol, a common functional group in small molecules, especially in small molecules prepared using diversity-oriented synthesis. This method comprises one element of a systematic approach to the target identification problem in chemical biology.
Wang
Xiang
X
Howard Hughes Medical Institute, Broad Institute of Harvard and MIT, 7 Cambridge Center, Cambridge, MA 02142, USA.
Imber
Brandon S
BS
Schreiber
Stuart L
SL
D000107:Acetylation; D000438:Alcohols / Q000737:chemistry; D002477:Cells / Q000737:chemistry* / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D017665:Hydroxyl Radical / Q000737:chemistry; D007093:Imidazoles / Q000737:chemistry; D007202:Indicators and Reagents / Q000737:chemistry*; D013058:Mass Spectrometry; D012685:Sepharose / Q000737:chemistry
D000438:Alcohols; D007093:Imidazoles; D007202:Indicators and Reagents; D017665:Hydroxyl Radical; C006900:N,N-carbonyldiimidazole; D012685:Sepharose