Abstract
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.
| Original language | English (US) |
|---|---|
| Title of host publication | Methods in Enzymology |
| Publisher | Academic Press Inc. |
| Pages | 83-107 |
| Number of pages | 25 |
| Edition | C |
| DOIs | |
| State | Published - Jan 1 2010 |
Publication series
| Name | Methods in Enzymology |
|---|---|
| Number | C |
| Volume | 481 |
| ISSN (Print) | 0076-6879 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
Cite this
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A practical guide to the use of monolayer purification and affinity grids. / Kelly, Deb; Dukovski, Danijela; Walz, Thomas.
Methods in Enzymology. C. ed. Academic Press Inc., 2010. p. 83-107 (Methods in Enzymology; Vol. 481, No. C).Research output: Chapter in Book/Report/Conference proceeding › Chapter
TY - CHAP
T1 - A practical guide to the use of monolayer purification and affinity grids
AU - Kelly, Deb
AU - Dukovski, Danijela
AU - Walz, Thomas
PY - 2010/1/1
Y1 - 2010/1/1
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=77957228396&partnerID=8YFLogxK
U2 - 10.1016/S0076-6879(10)81004-3
DO - 10.1016/S0076-6879(10)81004-3
M3 - Chapter
T3 - Methods in Enzymology
SP - 83
EP - 107
BT - Methods in Enzymology
PB - Academic Press Inc.
ER -