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Characterization of discrete oligomeric species of the regulatory protein RecA from Eschickz coli by Dynamic light scattering (DLS) (CAT#: STEM-MB-0551-WXH)
Introduction
RecA is a 38 kilodalton protein essential for the repair and maintenance of DNA. A RecA structural and functional homolog has been found in every species in which one has been seriously sought and serves as an archetype for this class of homologous DNA repair proteins. The homologous protein is called RAD51 in eukaryotes and RadA in archaea. RecA has multiple activities, all related to DNA repair. In the bacterial SOS response, it has a co-protease function in the autocatalytic cleavage of the LexA repressor and the λ repressor.
Principle
Dynamic Light Scattering (DLS) is an established and precise measurement technique for characterizing particle sizes in suspensions and emulsions. It is based on the Brownian motion of particles - this states that smaller particles move faster, while larger ones move slower in a liquid. The light scattered by particles contains information on the diffusion speed and thus on the size distribution.
The Dynamic Light Scattering (DLS) technique measures motion optically by recording the scattered light signal at a fixed angle. The particles are illuminated with a monochromatic, coherent light source (laser) and the light scattered by the particles is recorded.
The Dynamic Light Scattering (DLS) technique measures motion optically by recording the scattered light signal at a fixed angle. The particles are illuminated with a monochromatic, coherent light source (laser) and the light scattered by the particles is recorded.
Applications
DLS is used to characterize the size of various particles including proteins, polymers, micelles, Protein cages and virus-like particles, vesicles, carbohydrates, nanoparticles, biological cells, and gels.
Procedure
1. Sample preparation
2. Measurement by DLS instrument
3. Data analysis
2. Measurement by DLS instrument
3. Data analysis
Materials
• Dynamic Light Scattering (DLS) instruments (Photon Correlation Spectroscopy or Quasi-Elastic Light Scattering instruments)
• Dynamic Light Scattering Detector
• Dynamic Light Scattering Detector
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