The hydrodynamic radius of a lysozyme monomer was obtained from the Stokes-Einstein equation it is 18.6 for lysozyme chloride/water at concentrations from 0.43 to 3.08 mM (6.2-44.1 mg/mL), at the same pH and temperature. A Zimm-plot analysis provided the infinitely-dilute diffusion coefficient and the protein-concentration dependence of the diffusion coefficient. The diffusion coefficient of lysozyme was calculated as a function of protein concentration, salt concentration, temperature, and scattering angle. Turn off the machine, log your time of operation in the logbook.Dynamic light-scattering (DLS) studies are reported for lysozyme in aqueous magnesium chloride solutions at ionic strengths 0.6, 0.8, and 1.0 M for a temperature range 10-30 C at pH 4.0.Push start in the measurement window, collect at least 10 stable data points.Push start in the count rate window, monitor count rate, wait until it becomes stable, then stop it.On the computer program, go to "Tools" menu for experiment settings and optics configuration, based on what your experiment runs, you can adjust these parameters for best performance later.Put the cuvette frosty side left into the microsampler, close the lid. Inject the sample into the cuvette, but do not over fill.Remember there will be about 40µl deal volume inside the filter. To get your sample into the syringe, filter it through the microfilter.Rinse the microfilter with water to make sure there is no leakage. Unscrew to open the microfilter, add a 0.2µm filter membrane in between the 2 halves, and screw the microfilter tight. Typical applications of dynamic light scattering. Filter your sample whenever possible before applying it to the filter-kit. Dynamic light scattering (DLS), sometimes referred to as Quasi Elastic Light Scattering (QELS), is a non-invasive, well-established technique for measuring the size and size distribution of molecules and particles typically in the submicron region, and with the latest technology, lower than 1nm.If scattering is present, then the cuvette is not clean). Clean the cuvettes before and after use with water and ethanol dry the cell using "air-it." (It is a good idea to fill a cuvette with water and check for light scattering before testing your sample.Start the Dynamics program on the computer (icon on desktop). Turn on the measurement unit, then turn on the power unit, let it warm up for several minutes.Protocol for DynaPro99 Molecular Size Instrument (Dynamic Light Scattering) Measurements take only a few minutes and samples are generally recoverable.įor questions about getting started with a Dynamic Light Scattering project contact Dr. It measures the hydrodynamic radius down to 1.0nm and provides the distribution independent of solvent conditions so that the macromolecular behavior may be studied in virtually any solution condition. The DynaPro is highly sensitive in the detection of aggregation - even trace amounts - which often go undetected with other techniques such as gel filtration chromatography. The DynaPro-MS (MicroSampler) is an ultra-sensitive instrument combining 45 µl sample volumes with the highest sensitivity available over the 0.5 nm - 200 nm radius size range. Enhanced Protein Aggregation Detection Using Dual Angle Dynamic Light Scattering 04 November 2010 Contact sales Aggregates in protein formulations used for therapeutics have been shown to induce adverse reactions when administered. Protein Solutions DynaPro MSTC (temperature-controlled, micro-sampler)
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