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Study of Cu2+ binding to human serum albumin (HSA) by Dual polarization interferometry (DPI) (CAT#: STEM-MB-0395-WXH)

Introduction

Human serum albumin (HSA) is a high-molecular-weight endogenous plasma protein of 67 kDa. It is the main component of the blood transport system and reversibly binds a variety of endogenous (vitamins and lipids) and exogenous (drugs and toxins) molecules, distributing them to the target organs. HSA is also known to bind metal cations, which play crucial roles in human growth, development, cell division and synthesis of proteins and DNA. It is essential in the transport and metabolism of competitively bound Cu2+ ions in particular.




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 in human growth, development, cell division and synthesis of proteins and DNA.

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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