Journal: Drug Design, Development and Therapy
Article Title: Computational and nonglycosylated systems: a simpler approach for development of nanosized PEGylated proteins
Figure Lengend Snippet: SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with IPTG for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl β-D-1-thiogalactopyranoside.
Article Snippet: The expression of analogs was induced using 1 mM isopropyl β-d -1-thiogalactopyranoside (IPTG) (Sigma-Aldrich, Munich, Germany) and analyzed using 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) using Mini-PROTEIN® Tetra Cell system (Bio-Rad Laboratories, Hercules, CA, USA); the gel was developed using the Coomassie blue staining technique.
Techniques: SDS Page, Western Blot, Expressing, Marker, Transformation Assay, Positron Emission Tomography, Plasmid Preparation, Molecular Weight, Polyacrylamide Gel Electrophoresis