Study of Protein Translocation through the Membrane by Dual polarization interferometry (DPI) (CAT#: STEM-MB-0399-WXH)

Introduction

Translocation or transport of proteins and peptides across cell membrane into the cytosol is a complex process involving multistep mechanisms regulated by multicomponent translocation machineries. Many protein toxins from plants and microbial pathogens are able to translocate through the cell membrane and become active, exerting toxic effects by interacting with their counterpart molecules in the cytosol. A comprehensive analysis combining the structural characteristics of proteins and the physicochemical properties of the membrane have begun to unravel the underlying molecular mechanisms of various processes involved in toxin translocation across the cell membrane.

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Principle Applications Procedure Materials Notes Related Products

Principle

Dual polarization interferometry (DPI) is an analytical technique that allows the simultaneous determination of thickness, density, and mass of a biological layer on a sensing waveguide surface in real time. DPI focuses laser light into two waveguides. One of these functions as the "sensing" waveguide having an exposed surface while the second one functions to maintain a reference beam. A two-dimensional interference pattern is formed in the far field by combining the light passing through the two waveguides. The DPI technique rotates the polarization of the laser, to alternately excite two polarization modes of the waveguides. Measurement of the interferogram for both polarizations allows both the refractive index and the thickness of the adsorbed layer to be calculated. These measurements can be used to infer conformational information about the molecular interactions taking place, as the molecule size (from the layer thickness) and the fold density (from the RI) change.

Applications

Study of Protein Translocation through the Membrane

Procedure

1. Setting of dual polarization interferometry
2. Preparing the DPI sensor chip
3. Immobilization of target on DPI biosensor
4. Reagent was injected to react
5. Quantitative analysis

Materials

• DPI biosensor
• DPI sensor chip

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