The dried lipid film was re-dissolved in diethyl ether followed by addition of HB150 buffer (150 mM KCl, 20 mM Hepes (pH 7.5)) containing 1 mM DTT. After dispersal by sonication, the diethyl ether was removed by evaporation. Liposomes were extruded using polycarbonate membranes with ...
U2OS cells carrying latent HHV-6A treated with solvent control or TSA were lysed in RIPA buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 1% Triton X-100, 1% NP-40, 0.1% SDS, 10% Glycerol and protease inhibitor cocktail (Roche)). Protein lysates were incubated with control...
Cell were resuspended in Buffer A containing 1 mm EDTA, 0.5 μg/ml lysozyme, phenylmethylsulfonyl fluoride. Cell lysis was performed by a combination of three freeze and thawing cycles and sonication. Lysate was centrifuged at 16,000 x G for 30 min and then dialyzed against 50 mm Tris, ...
The recombinant protein was expressed in SoluBL21 (Genlantis) competent cells hosting pRARE plasmids by following the manufacturer's protocol, then extracted with 20 mM Tris–HCl lysis buffer (pH 7.5), purified by nickel affinity chromatography (Ni Sepharose 6 Fast Flow, GE Healthcare), and ...
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of COVID-19, generates multiple protein-coding, subgenomic RNAs (sgRNAs) from a longer genomic RNA, all bearing identical termini with poorly understood roles in regulating
Cells from 5-day cultures were har- vested from solid YM media dissolved in 1% PBS buffer, filtered through 30 μm filter (Partec GmbH, Germany) and inoculated in the artificial nectar medium (20 cells per μL = 3000 cells per well) using BD FACSAria III cell sorter (BD Biosciences, ...