reconstitution buffer  (Thermo Fisher)

Journal: Life Science Alliance

Article Title: Cryo-EM structures reveal the H + /citrate symport mechanism of Drosophila INDY

doi: 10.26508/lsa.202402992

Figure Lengend Snippet: The SDS–PAGE results confirm similar levels of protein reconstitution in each liposome preparation. The gel is one representative result from at least two independent experiments.

Article Snippet: The dried lipids were resuspended in reconstitution buffer containing 20 mM MES-NaOH pH 6.0, and 50 mM NaCl for [ 14 C]-citrate transport assay, or 20 mM MES-NaOH pH 6.0, 150 mM KCl, and 1 mM pyranine (trisodium 8-hydroxypyrene-1,3,6-trisulfonate; MedChemExpress) for H + transport assay.

Techniques: SDS Page

Journal: Scientific Reports

Article Title: Electromagnetic waves destabilize the SARS-CoV-2 Spike protein and reduce SARS-CoV-2 Virus-Like particle (SC2-VLP) infectivity

doi: 10.1038/s41598-025-01896-1

Figure Lengend Snippet: Electromagnetic waves reduce infectivity of SARS-CoV-2 Virus-Like Particles (SC2-VLPs). (A) Diagram of SC2-VLP production, electromagnetic wave treatment and infectivity assay. Plasmids expressing each of the SARS-CoV-2 structural proteins (Nucleocapsid (N); Matrix (M); Envelope (E); and Spike (S)), or a plasmid encoding a packaging signal and luciferase transcript (Luc-PS9) were transfected into viral producer cells (HEK293T). SC2-VLPs were then collected, left unexposed or exposed to various electromagnetic wave frequencies (1–6 GHz) and used to infect target cells (HEK293T-ACE2-TMPRSS2). The next day, luciferase was measured as a readout of infectivity. Created in https://BioRender.com . (B) Western blot of pelleted Spike (S), Nucleocapsid (N), Membrane (M), Envelope (E) in unexposed virus particles. The image is representative of two independent experiments. (C) SC2-VLP infectivity assay. Frequencies (1–6 GHz) were tested over a range of exposure times (2–10 min). Data represent compiled mean +/- SD of 5 independent experiments. The luciferase signal for each frequency/exposure time was performed in triplicate. P-values indicate Wilcoxon matched-pair signed rank test of unexposed compared to exposed samples. ( n = 90 (Unexposed); n = 36 (1.0–2.5 GHz & 2.5–3.5 GHz) n = 15 (3.5–4.8 GHz), n = 33 (4.8–6 GHz), n = 18 (MW)). RLU: Renilla Luciferase Units (RLU). MW: Microwave.

Article Snippet: The cells were then transferred to an opaque white 96-well flat-bottom plate, mixed with 50 μL reconstituted luciferase assay buffer (Promega), and luciferase measured immediately on a plate reader (Biotek Synergy H1).

Techniques: Infection, Virus, Expressing, Plasmid Preparation, Luciferase, Transfection, Western Blot, Membrane