Volume 10 Issue 5, May 2015

This protocol uses fluorescence recovery after photobleaching (FRAP) to dissect the reaction dynamics leading to protein turnover in macromolecular complexes in living cells.

This protocol uses RNAi-based biosynthetic pathway screens combined with genetic and biochemical analyses to identify the functions of non-nucleic acid-based metabolites such as lipids beyond their roles in metabolism.

Thanks to advances in MS/MS acquisition rates and improvements in sample preparation and HPLC separation, it is now possible to analyze the yeast proteome in a single 70-min LC-MS/MS run.

This protocol describes a transcriptome-wide approach to detect adenosine-to-inosine editing sites in RNAs using inosine chemical erasing combined with deep sequencing (ICE-seq).

This protocol describes the FTMap family of web servers for determining and characterizing ligand-binding hot spots of macromolecules, including FTSite for predicting ligand-binding sites, FTFlex for accounting for side chain flexibility, FTMap/param for parameterizing additional probes, and FTDyn for mapping ensembles of protein structures.

This protocol allows the identification of AMPylated and auto-AMPylated proteins using high-density protein microarrays fabricated using NAPPA technology. Target protein modification is determined via copper-catalyzed azide-alkyne cycloaddition.

This protocol enables collection of airborne particulate matter; and after sample pretreatment, it allows sufficient quantities of microbial DNA to be extracted and prepared for downstream applications such as metagenomic sequencing.

Site-specific labeling of proteins with small fluorophores is advantageous for imaging. Lemke et al. describe how to site-specifically label membrane proteins with organic fluorophores by incorporating non-canonical amino acids via Amber suppression technology.

Moro and colleagues describe improved methods for tissue-specific isolation and analysis of mouse ILC2s from fat, lung, bronchoalveolar lavage fluid and small intestine tissue.

This protocol describes the design of a cyclic herpes simplex 1 virus-thymidine kinase reporter and its use in PET imaging of chemotherapy-induced apoptosis in cancer cells in vitro and in vivo.