pet sac abeta  (Addgene inc)

Journal: The FEBS journal

Article Title: Membrane protein chaperone and sodium chloride modulate the kinetics and morphology of amyloid beta aggregation.

doi: 10.1111/febs.16967

Figure Lengend Snippet: Fig. 1. Concentration of NaCl affects the kinetics of protein aggregation. (A) Time-dependent ThT fluorescence measurements of 10 μM Aβ42 (Peptide: ThT ratio = 1 : 1) incubated at 37 °C in the absence (red) or presence of NaCl at 6.25 mM (cyan), 12.5 mM (green), 25 mM (blue), 50 mM (gray), 100 mM (purple), 150 mM (yellow), and 200 mM (orange). All the kinetic experiments were performed in a 37 °C fluo- rescence plate reader under shaking conditions of 250 rpm and fluorescence data were collected every 10 min. Each data point represents the mean value and error bars represent standard error with n = 3. (B) Aβ42 aggregation kinetics were fitted to a sigmoidal function and the aggregation lag time was plotted as a function of NaCl concentration. Each data point represents the mean value and error bars represent standard error with n = 3.

Article Snippet: Aβ40 and Aβ42 plasmids [pET-Sac-Abeta (M1-42) and pET-Sac-Abeta (M1-40); Addgene plasmids #71875 and #71876, respectively] containing exogenous methionine at the N-terminus were gifts from Dominic Walsh [80].

Techniques: Concentration Assay, Incubation

Journal: The FEBS journal

Article Title: Membrane protein chaperone and sodium chloride modulate the kinetics and morphology of amyloid beta aggregation.

doi: 10.1111/febs.16967

Figure Lengend Snippet: Fig. 2. Electrostatic repulsion of Aβ42 affects the formation of protein aggregates. (A) Primary sequence of Aβ40 and Aβ42 with the charged residues highlighted. Additional hydrophobic residues in Aβ42 are highlighted in red. (B, C) Electrostatic surface potential (B) and ribbon diagram (C) of the Aβ42 fibril from the Protein Data Bank (PDB: 2NAO) visualized in PYMOL. Positively and negatively charged side chains of amino acids are shown in blue and red, respectively. Hydrophobic residues are shown in white and the protein backbone in green. Electrostatic repulsions are highlighted between fibril layers.

Article Snippet: Aβ40 and Aβ42 plasmids [pET-Sac-Abeta (M1-42) and pET-Sac-Abeta (M1-40); Addgene plasmids #71875 and #71876, respectively] containing exogenous methionine at the N-terminus were gifts from Dominic Walsh [80].

Techniques: Sequencing

Journal: The FEBS journal

Article Title: Membrane protein chaperone and sodium chloride modulate the kinetics and morphology of amyloid beta aggregation.

doi: 10.1111/febs.16967

Figure Lengend Snippet: Fig. 6. TEM and SEM reveal that cpSRP43 can change the morphology of Aβ40 and Aβ42 aggregates. (A, B) Aβ40 (20 μM) monomer was incubated at 37 °C for 3 days either in the absence (A) or presence (B) of 10-fold excess cpSRP43. For each sample, 1 μL was dried and visualized by TEM. The images in both (A) and (B) were obtained at a magnification of 15K×. Scale bars are 500 nm. (C, D) Aβ42 monomer (10 μM) was incubated with 1 μM Aβ42 seeds in the absence (C) or presence (D) of 10-fold excess cpSRP43 for 4 days at 37 °C. For each sample, 1 μL was dried and visualized by SEM. The images in both (C) and (D) were captured at 600×. Scale bars are 50 μm. Data are rep- resentative of three independent experiments.

Article Snippet: Aβ40 and Aβ42 plasmids [pET-Sac-Abeta (M1-42) and pET-Sac-Abeta (M1-40); Addgene plasmids #71875 and #71876, respectively] containing exogenous methionine at the N-terminus were gifts from Dominic Walsh [80].

Techniques: Incubation