Author: Jacob W. Myerson; Priyal N. Patel; Nahal Habibi; Landis R. Walsh; Yi-Wei Lee; David C. Luther; Laura T. Ferguson; Michael H. Zaleski; Marco E. Zamora; Oscar A. Marcos-Contreras; Patrick M. Glassman; Ian Johnston; Elizabeth D. Hood; Tea Shuvaeva; Jason V. Gregory; Raisa Y. Kiseleva; Jia Nong; Kathryn M. Rubey; Colin F. Greineder; Samir Mitragotri; George S. Worthen; Vincent M. Rotello; Joerg Lahann; Vladimir R. Muzykantov; Jacob S. Brenner
Title: Supramolecular Organization Predicts Protein Nanoparticle Delivery to Neutrophils for Acute Lung Inflammation Diagnosis and Treatment Document date: 2020_4_18
ID: ezrkg0dc_74
Snippet: Glutamic acid residues (E20-tag) were inserted at the C-terminus of enhanced green fluorescent protein (EGFP) through restriction cloning and site-directed mutagenesis as previously reported. 64 Proteins were expressed in an E. coli BL21 strain using standard protein expression protocol. Briefly, protein expression was carried out in 2xYT media with an induction condition of 1 mM IPTG and 18 °C for 16 h. At this point, the cells were harvested, .....
Document: Glutamic acid residues (E20-tag) were inserted at the C-terminus of enhanced green fluorescent protein (EGFP) through restriction cloning and site-directed mutagenesis as previously reported. 64 Proteins were expressed in an E. coli BL21 strain using standard protein expression protocol. Briefly, protein expression was carried out in 2xYT media with an induction condition of 1 mM IPTG and 18 °C for 16 h. At this point, the cells were harvested, and the pellets were lysed using 1% Triton-X-100 (30 min, 37 °C)/DNase-I treatment (10 minutes). Proteins were purified using HisPur cobalt columns. After elution, proteins were preserved in PBS buffer. The purity of native proteins was determined using 8% SDS−PAGE gel.
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