Laboratory Methods and Program Resources

Laboratory Methods

The Structural Biology program has transitioned from Common Fund support. For more information, please visit Program Snapshot page.

Please note that since the Structural Biology program is no longer supported by the Common Fund, the program website is being maintained as an archive and will not be updated on a regular basis.

The Common Fund supported Structural Biology Program has funded the Joint Center for Innovative Membrane Protein Technologies-Complexes (JCIMPT-Complexes) and the Membrane Protein Expression Center (MPEC). These centers are research networks comprised of multiple integrated projects to develop new technologies and methods to enable the production of large amounts of homogeneous membrane proteins and determine protein structure. ​

Experimental Protocols

• Viral Stock generation
• Cell free Protein Production
• Membrane Prep
• Micro-Purification
• Large-Scale Protein Purification
• Preparation of Cholesterol in Detergent
• Measure Cell Surface Expression
• Microcoil NMR<
• IMP Refolding for NMR Studies

Video Tutorial

• Crystallizing membrane proteins
• Other videos

Experimental Protocols

Techniques for membrane protein research:

• A general protocol for the crystallization of membrane proteins for X-ray structural investigation
• Selecting optimum eukaryotic integral membrane proteins for structure determination by rapid expression and solubilization screening
• Cell-free complements in vivo expression of E. coli membrane proteome
• A green fluorescent protein screen for identification of well-expressed membrane proteins from a cohort of extremophilic organisms
• A high throughput assay for membrane protein stabilization by ligands, buffers, detergents, lipids; and high quality membrane proteins in S. cerevisciae
  • Overexpression and purification of integral membrane proteins in yeast
  • Selecting optimum eukaryotic integral membrane proteins for structure determination by rapid expression and solubilization screening
  • Ratiocinative screen of eukaryotic integral membrane protein expression and solubilization for structure determination
  • A survey of integral alpha-helical membrane proteins
• Efficient expression screening of human membrane protein in transiently transfected Human Embryonic Kidney 293S cells
• Overexpressing human membrane proteins in stably transfected and clonal human embryonic kidney 293S cells

Expression systems used at MPEC.