A PRACTICAL GUIDE TO THE USE OF MONOLAYER PURIFICATION AND AFFINITY GRIDS

作者：Kelly Deborah F^*; Dukovski Danijela; Walz Thomas

来源：METHODS IN ENZYMOLOGY, VOL 481: CRYO-EM, PART A - SAMPLE PREPARATION AND DATA COLLECTION , ELSEVIER ACADEMIC PRESS INC, 525 B STREET, SUITE 1900, SAN DIEGO, CA 92101-4495 USA, 83-107, 2010.

DOI：10.1016/S0076-6879(10)81004-3

Lipid monolayers have traditionally been used in electron microscopy (EM) to form two-dimensional (2D) protein arrays for structural studies by electron crystallography. More recently, monolayers containing Nickel-nitrilotriacetic acid (Ni-NTA) lipids have been used to combine the purification and preparation of single-particle EM specimens of His-tagged proteins into a single, convenient step. This monolayer purification technique was further simplified by introducing the Affinity Grid, an EM grid that features a predeposited Ni-NTA lipid-containing monolayer. In this contribution, we provide a detailed description for the use of monolayer purification and Affinity Grids, discuss their advantages and limitations, and present examples to illustrate specific applications of the methods.

• 出版日期2010