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. Author manuscript; available in PMC: 2011 Mar 23.
Published in final edited form as: Apoptosis. 2010 Dec;15(12):1444–1452. doi: 10.1007/s10495-010-0544-2

Figure 4. The solubility of RAIDD is sensitive to salt concentration.

Figure 4

A. Solubility assay of salt concentration effect on RAIDD aggregation. RAIDD protein was prepared 1mg/ml concentration in buffer containing 20 mM Tris-HCl pH 8.0, 150 mM NaCl. Turbidity of each samples incubated with various different salt concentration for 3 days at 4°C, as measured by the optical density at 600 nm. Values are means SD of n=3. B. Proportion of aggregated full-length RAIDD following incubation with various NaCl concentration at 4°C for 3 days after centrifugation expressed as a percentage of the amount of initial concentration of RAIDD. Protein concentration of soluble portion of RAIDD in the solution was checked by Bradford assay. Values are means SD of n=3. The soluble portion of RAIDD in the solution was also loaded onto SDS-PAGE. Protein bands on a 15 % SDS-PAGE gel were detected by coomasie blue staining. The migration of molecular size marker is indicated. C. Solubility assay of salt effect on reversible RAIDD aggregation. Prepared RAIDD protein was incubated at R.T. for 12 hours and let protein aggregate. The aggregate was placed in the buffer containing various salt concentration indicated for a day and turbidity of each samples was measured by the optical density at 600 nm. Values are means SD of n=3. D. Proportion of reduced aggregate following incubation with various NaCl concentrations for a day expressed as a percentage of the amount of initial concentration of RAIDD. Protein concentration of solubilized portion of RAIDD in the solution was checked by Bradford assay. Values are means SD of n=3.