Proteomic Analysis of Scaffold Proteins in the ERK Cascade
ERK cascade scaffolds serve as docking platforms to coordinate the assembly of multiprotein complexes that contribute to the spatial and temporal control of ERK signaling. Given that protein–protein interactions are essential for scaffold function, determining the full repertoire of scaffold binding partners will likely provide new insight into the regulation and activities of the ERK cascade scaffolds. In this chapter, we describe methods to identify scaffold interacting proteins using a proteomics approach. This protocol is based on the affinity purification of scaffold complexes from tissue culture cells and utilizes mass spectrometry to identify the protein constituents of the complex.
- Isolation and Culture of Mouse Pancreatic Islets for Ex Vivo Imaging Studies with Trappable or Recombinant Fluorescent Probes
- Transfer of Eukaryote Suspension Cultures
- Isolation and Differentiation of Medaka Embryonic Stem Cells
- 红细胞膜的制备及其成份分析(磷脂、胆固醇)
- A pH-Indicator-Based Screen for Hydrolytic Haloalkane Dehalogenase
- Standardized Isolation of Human Mesenchymal Stromal Cells with Red Blood Cell Lysis
- 自由基(free radical)
- The Use of RAPD for Isolate Identification of Arbuscular Mycorrhizal Fungi
- Taq Polymerase (EC 2.7.7.7): With Particular Emphasis on Its Use in PCR Protocols
- 酵母遗传学方法9:酵母免疫荧光