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. Author manuscript; available in PMC: 2017 Feb 15.
Published in final edited form as: Langmuir. 2016 Jan 4;32(2):551–559. doi: 10.1021/acs.langmuir.5b03445

Figure 1.

Figure 1

Conceptual diagram showing stabilized monolayer affinity grid fabrication and utilization. (Top left) A fluid-phase mixed lipid monolayer comprised of Ni2+:NTA-PEG2000-DSPE and mPEG350-DTPE (1:99 or 5:95 mol:mol) was compressed until the mPEG350-DTPE component entered the brush regime and the diyne lipid chains were fully condensed; (Top center) the condensed lipid monolayer film was deposited onto carbon-coated TEM grids via Langmuir-Schaefer (LS) transfer; (Top right) photopolymerization of the LS film, followed by Ni2+ activation, produces a grid with a stable affinity capture coating; (Bottom right) solutions containing the His-tag protein of interest are deposited onto the coated grids, either as a clarified cell lysate or as a purified protein sample; (Bottom center) blotting and rinsing of the sample removes non-target material prior to (Bottom left) cryofixation (or negative staining) and sample imaging.