This protocol describes the methods to prepare and isolate the vesicles, equipment to observe them under temperature-controlled conditions and three examples of fluorescence analysis: (i) fluorescence spectroscopy with an environment-sensitive dye (laurdan); (ii) two-photon microscopy of the same dye; and (iii) quantitative confocal microscopy to determine component partitioning between raft and nonraft phases. TY - JOURT1 - Elucidating membrane structure and protein behavior using giant plasma membrane vesicles AU - Sezgin, Erdinc AU - Kaiser, Hermann Josef AU - Baumgart, Tobias AU - Schwille, Petra AU - Simons, Kai AU - Levental, Ilya PY - 2012/6Y1 - 2012/6N2 - The observation of phase separation in intact plasma membranes isolated from live cells is a breakthrough for research into eukaryotic membrane lateral heterogeneity, specifically in the context of membrane rafts.
GPMV preparation and isolation, including fluorescent labeling and observation, can be accomplished within 4 h. These observations are made in giant plasma membrane vesicles (GPMVs), which can be isolated by chemical vesiculants from a variety of cell types and microscopically observed using basic reagents and equipment available in any cell biology laboratory.
By using UPLC separation, the CNP product was well separated into ten fractions within 4.0 min.
Based on high-accuracy MS and MS/MS analyses, the CNP species were revealed to display six kinds of chemical formulas, including (C.
Copper is one of the most abundant biological metals, and its chemical properties mean that organisms need sophisticated and multilayer mechanisms in place to maintain homoeostasis and avoid deleterious effects.
Studying copper proteins requires multiple techniques, but electron paramagnetic resonance (EPR) plays a key role in understanding Cu(II) sites in proteins.
These observations are made in giant plasma membrane vesicles (GPMVs), which can be isolated by chemical vesiculants from a variety of cell types and microscopically observed using basic reagents and equipment available in any cell biology laboratory.
Microscopic phase separation is detectable by fluorescent labeling, followed by cooling of the membranes below their miscibility phase transition temperature.
This chapter discusses the origins of curcumin’s biological activities in light of its structure-activity relationship.
The structure of curcumin is comprised of the central 1,6-heptadiene-3,5-dione bearing two terminal phenolic rings.
A fast and accurate ultra-performance liquid chromatography coupled with electrospray ionisation quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-Q-TOF-MS/MS) method was developed for the separation and structural elucidation of fluorescent carbon nanoparticles (CNP).