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Fig. 4 | Biological Procedures Online

Fig. 4

From: Characterising protein/detergent complexes by triple-detection size-exclusion chromatography

Fig. 4

SEC as monitored by static light scattering of Mistic solubilised in different alkyl maltosides in the (a) absence and (b) presence of 6 M urea. a Excess Rayleigh ratios at 90° for Mistic solubilised in DDM, UM, DM, and NM are depicted as functions of elution volume. In the absence of urea, the traces of NM and DM show two peaks, indicating separation of PDCs and protein-free micelles. Single peaks for UM and DDM indicate co-elution of PDCs and micelles. b At 6 M urea, for all detergents except DDM, unfolded protein and detergent micelles are separated. The elution volume of the protein is at ~9.3 mL for all measurements, whereas the elution volumes of detergent micelles change according to their size. For DDM, unfolded detergent-free Mistic and DDM micelles co-elute [11]. Additional peaks in the SLS signal at ~8.5 mL in the DDM and UM traces are so-called “system peaks”; these peaks are typically caused by injected air or shedding of the SEC column and do not show up in UV and RI signals. 1 mg/mL Mistic, 50 mM Tris, pH 7.4, 50 mM NaCl, c det,sample = CMC8 M urea + 5 mM, c det,buffer = CMCbuffer + 2 mM, flow rate 0.4 mL/min, RT

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