MOBILITY STUDY OF ALLYLAMINE BY FRAP

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are interested to study the mobility of surface adsorbed polyallylamine using Fluorescence Recovery After Photobleaching (FRAP), a single molecule based technique. This technique gives us a direct measure of the rates of lateral transport of proteins and lipids in cell membranes. For us, the three issues of greatest interest are: (1) Does poly(allylamine) come off of a layer-by-layer surface [with an underlayer of poly(styrenesulfonate)] under any conditions; e.g., acid, base high salt concentration, (2) is there any lateral mobility of the outer poly(allyamine) layer under any conditions; i.e., FRAP within this outer layer, and (3) how does our novel covalently-cross-linked poly(allylamine) surface compare with conventional ionically-cross-linked, layer-by-layer surfaces in terms of stability and lateral mobility. For this the polyallylamine is labeled with a fluorescent dye that can be excited by the 514 nm line of an Argon-ion laser which in turn is coupled to a confocal microscope.