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Proteins were extracted from cells using a lysate buffer (Intron Biotechnology, Gyeonggi-Do, Korea) and quantified by the Bradford assay. Aliquots containing of 20 μg protein were electrophoresed on 10% SDS-PAGE gels for 2 hours at 80 V. Proteins were transferred to PVDF membranes (Bio Rad) for 1 hour at 15 V using a Semi-Dry transfer unit (Bio Rad). Following preincubation with a blocking agent, the membranes were incubated with a monoclonal antibody to clusterin (Millipore, Temecula, CA, USA), followed by incubation with species specific secondary antibody. As a loading control, the membranes were incubated with a monoclonal antibody to β-actin (Cell Signaling, Danver, MA USA). Protein expression was developed using ECL (Enhanced chemiluminescence; Thermo Fisher Scientific) and analyzed with the Quantity One program (Bio Rad.). million smoked daily.. development of many multigene prognostic tests to improve assessment. Cultivation in suspension allows up-scaling of the photo bioreactors. A prospective study of a curated sample of head CT's were performed in an emergency department. Raters independently reviewed the scans. Perpendicular maximal dimensions (A and B) were measured on an axial CT image. The ‘C’ dimension was a product of slice thickness and number of slices with visible bleed..

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