In situ detection of the eIF4F translation initiation complex in mammalian cells and tissues

The eukaryotic translation initiation complex eIF4F plays an important role in gene expression. The methods that are used to monitor the formation of the eIF4F complex are usually indirect and provide no information on its subcellular localization. This protocol describes a proximity ligation assay-based procedure allowing the direct in situ visualization of the eIF4F complex, as well as its abso-lute quantification per cell using adapted image analysis software.For complete details on the use and execution of this protocol, please refer to Boussemart et al. (2014).

Automated summary

This protocol describes a procedure utilizing proximity ligation assay for direct visualization and absolute quantification of the eIF4F complex per cell, allowing for detailed analysis of its subcellular localization.