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Molecular mass analysis of OmpF porin by Static light scattering (SLS) (CAT#: STEM-MB-0572-WXH)

Introduction

OmpF porin, an integral membrane protein that serves as a passive pore for the passage of small hydrophilic molecules across the outer membrane of Escherichia coli, has been shown to have virtually identical structures in two completely different (trigonal and tetragonal) crystal forms. OmpF protein is one of the best studied membrane proteins up to now. The mature protein is comprised of 340 amino acids, and forms aqueous, voltage-gated channels that span the outer membrane and allow the diffusion of small polar molecules.




Principle

Static light scattering is a technique in physical chemistry that measures the intensity of the scattered light to obtain the average molecular weight Mw of a macromolecule like a polymer or a protein in solution. Measurement of the scattering intensity at many angles allows calculation of the root mean square radius, also called the radius of gyration Rg. By measuring the scattering intensity for many samples of various concentrations, the second virial coefficient, A2, can be calculated.

Applications

The main applications of static light scattering is molecular mass determination of macromolecules, such as proteins and polymers, as it is possible to measure the molecular mass of proteins without any assumption about their shape.

Procedure

1. Sample preparation
2. Measurement by SLS instrument
3. Data analysis

Materials

• Right-Angle Light Scattering (RALS) Detector
• Low-Angle Light Scattering (LALS) Detector
• Hybrid RALS/LALS Detector
• Multi-Angle Light Scattering (MALS) Detector