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How to small single-domain proteins fold? Fold
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Protein solutions (protein concentrations: 7 µg/ml to 1 mg/ml) were tilled into quartz-cuvettes with an optical pathlength of 1 cm (104F-QS, Hellma, Muehlheim, Germany)
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The lowest protein concentration of 7 µg/ml reveals a large red shift with λ^sub max^ of ~345 nm, whereas protein concentrations of 50 µg/ml and 1 mg/ml result in λ^sub max^-values ~340 nm for the unfolded states
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The final water content was measured by weighing shortly before closing the sample holders and was 0
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Acetone and Isopar solvents are miscible, but methanol and Isopar solvents are not
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These fractions were plotted versus Mg concentration and fitted to the cooperative binding equation F = F^sup max^([Mg]^sup n^/([Mg]^sup n^ + (K^sup Mg^^sub D^)^sup n^)), to yield an apparent dissociation constant K^sup Mg^^sub D^for Mg^sup 2+^, and an apparent cooperativity coefficient, n
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