After LPS binding, membranes were washed 3 times, 10 min each time in the above buffers and visualized for radioactivity on Bas 2000 radioimaging system (Fuji, Japan).
For LPS binding, membranes were then blocked by 2 wt% BSA in PBS, pH 7.4, for 1 h at RT and subsequently incubated with radiolabelled LPS (40 µg/mL; 0.13×106 cpm/µg) for 1 h at RT in PBS [32].
To avoid non-specific binding, membranes were blocked with 5% nonfat milk protein in PBS/Tween at RT for 1 hour.
In contrast to DNA, which offers sequence-specific binding, membrane is less defined, although head group-specific binding and membrane curvature provide specificity in some cases that distinguish between different membranes in cells.
The regulation of MGD1, the committing enzyme of galactolipid biosynthesis in Arabidopsis, via feedback regulatory loops and control of protein binding to membranes, is also detailed.
We show that SBD binding to membranes is dependent on the presence of cholesterol, sphingomyelin, and complex glycolipids.
We conclude that membrane binding is a necessary function of the Imp2 F-BAR domain in cytokinesis to form an efficient scaffold for its partners.
Proteins were electrophoretically transferred onto PVDF membranes, and after overnight exposure to 3% BSA to block nonspecific binding, the membranes were incubated with a polyclonal FGF21 antibody (Santa Cruz Biotechnology Inc, 1 1000).
The binding of glycan binding proteins to glycan receptors on cell membranes is fundamentally a kinetic process.
A general prerequisite for a fusion protein/target membrane binding assay is a soluble and correctly oligomeric form of the viral fusion protein ectodomain.
