Optimized negative staining: a high-throughput protocol for examining small and asymmetric protein structure by electron microscopy.

ABSTRACT: Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of natural proteins have a molecular mass between 40 - 200 kDa, a robust and high-throughput method with a nanometer resolution capability is needed. Negative staining (NS) electron microscopy (EM) is an easy, rapid, and qualitative approach which has frequently been used in research laboratories to examine protein structure and protein-protein interactions. Unfortunately, conventional NS protocols often generate structural artifacts on proteins, especially with lipoproteins that usually form presenting rouleaux artifacts. By using images of lipoproteins from cryo-electron microscopy (cryo-EM) as a standard, the key parameters in NS specimen pr

SUBMITTER: Rames M 

PROVIDER: S-EPMC4710468 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature