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Published byMonica Webb Modified over 9 years ago
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CIRCULAR DICHROISM SPECTROSCOPY structural analysis of nmol samples of macromolecules Prof. Eric Wickstrom
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Circular Dichroism is the difference in absorption between left and right hand circularly polarised light in chiral molecules. A chiral molecule is one with a low degree of symmetry which can exist in two mirror image isomers. Illustrated above is an example of circular dichroism in glucose, a simple sugar.
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orthogonal electronic (E) and magnetic (B) components of linearly polarized light EE BB
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electronic field of linearly polarized light (left) right-handed circularly polarized light (right)
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Ellipticity,
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Prism Polarizer
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The most commonly used units are mean residue ellipticity, (degree·cm 2 /dmol), and the difference in molar extinction coefficients called the molar circular dichroism, ε L -ε R =Δε (liter/mol·cm). The molar ellipticity [ ] is related to the difference in extinction coefficients by [ ] = 3298 Δε.
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Typical Initial Concentrations Protein Concentration: 0.5 mg/ml Cell Path Length: 0.5 mm Stabilizers (Metal ions, etc.): minimum Buffer Concentration : 5 mM or as low as possible while maintaining protein stability Contaminants: Unfolded protein, peptides, particulate matter (scattering particles)
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