Study of Interaction of Nisin with Planar Lipid Bilayers by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0046-WXH)

Name: Study of Interaction of Nisin with Planar Lipid Bilayers by Fluorescence recovery after photobleaching (FRAP)
SKU: STEM-MT-0046-WXH
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Introduction

Nisin, a prominent member of the lantibiotic family of antimicrobial agents, has wide application as a food preservative despite poor understanding of its mode of action.
Nisin associates with planar lipid bilayers in the absence of an applied membrane potential causing an array of effects consistent with adsorption of nisin onto the membrane surface which involves inhibition of the lateral diffusion and fluorescence of the lipid probe N-(7--1,2,3-benzoxadiazol-4-yl) phosphatidylethanolamine (NBD-PE) and a reduction of the capacitance of the bilayer. Nisin adsorption is dependent on phospholipid composition.

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Principle Applications Procedure Materials

Principle

Fluorescence recovery after photobleaching (FRAP) is a microscopy technique capable of quantifying the mobility of molecules within cells. By exploiting the phenomenon of photobleaching, fluorescent mole- cules within a region of interest can be selectively and irreversibly 'turned off'. It is capable of quantifying the two-dimensional lateral diffusion of a molecularly thin film containing fluorescently labeled probes, or to examine single cells.

Applications

• Characterization of the mobility of individual lipid molecules within a cell membrane.
• Analysis of molecule diffusion within the cell
• Study of protein interaction partners, organelle continuity and protein trafficking.

Procedure

1. An initial fluorescence of fluorescent molecules is measured in the region of interest (ROI).
2. The fluorescent molecules are rapidly photobleached by focusing the high-intensity laser beam onto the defined area.
3. The exchange of bleached molecules with unbleached molecules from the surrounding region is followed over time using a low-intensity laser.