2 articles published in JoVE
Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy Ramakrishnan B. Kumar1, Lin Zhu2, Hans Hebert1,2, Caroline Jegerschöld1,2 1Department of Biosciences and Nutrition, Karolinska Institutet, 2School of Technology and Health, KTH Royal Institute of Technology Many proteins perform their function when attached to membrane surfaces. The binding of extrinsic proteins on nanodisc membranes can be indirectly imaged by transmission electron microscopy. We show that the characteristic stacking (rouleau) of nanodiscs induced by the negative stain sodium phosphotungstate is prevented by the binding of extrinsic protein.
Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag Johan Nilvebrant1, Tove Alm1, Sophia Hober1 1School of Biotechnology, Department of Proteomics, Royal Institute of Technology A novel and highly efficient two-step affinity chromatography protocol has been developed and is described in detail. The method is based on a small purification tag with two inherent affinities and is applicable to a wide range of target proteins with different properties.