A practical guide to the use of monolayer purification and affinity grids

Deb Kelly, Danijela Dukovski, Thomas Walz

Research output: Chapter in Book/Report/Conference proceedingChapter

16 Citations (Scopus)

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 languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages83-107
Number of pages25
EditionC
DOIs
StatePublished - Jan 1 2010

Publication series

NameMethods in Enzymology
NumberC
Volume481
ISSN (Print)0076-6879

Fingerprint

Purification
Monolayers
Electron Microscopy
Lipids
Electron microscopy
Protein Array Analysis
Crystallography
Electrons
Proteins
nickel nitrilotriacetic acid

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Kelly, D., Dukovski, D., & Walz, T. (2010). A practical guide to the use of monolayer purification and affinity grids. In Methods in Enzymology (C ed., pp. 83-107). (Methods in Enzymology; Vol. 481, No. C). Academic Press Inc.. https://doi.org/10.1016/S0076-6879(10)81004-3
Kelly, Deb ; Dukovski, Danijela ; Walz, Thomas. / A practical guide to the use of monolayer purification and affinity grids. Methods in Enzymology. C. ed. Academic Press Inc., 2010. pp. 83-107 (Methods in Enzymology; C).
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Kelly, D, Dukovski, D & Walz, T 2010, A practical guide to the use of monolayer purification and affinity grids. in Methods in Enzymology. C edn, Methods in Enzymology, no. C, vol. 481, Academic Press Inc., pp. 83-107. https://doi.org/10.1016/S0076-6879(10)81004-3

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 proceedingChapter

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Kelly D, Dukovski D, Walz T. A practical guide to the use of monolayer purification and affinity grids. In Methods in Enzymology. C ed. Academic Press Inc. 2010. p. 83-107. (Methods in Enzymology; C). https://doi.org/10.1016/S0076-6879(10)81004-3