Journal: iScience

Article Title: Dry molten globule conformational state of CYP11A1 (SCC) regulates the first step of steroidogenesis in the mitochondrial matrix

doi: 10.1016/j.isci.2024.110039

Figure Lengend Snippet:

Article Snippet: Lubrol , MP Biomedicals , 02195299-CF.

Techniques: Protease Inhibitor, RIA Assay, Plasmid Preparation, Expressing, Isolation, Lysis

Journal: bioRxiv

Article Title: Sphingolipids with Very Long-chain Fatty Acids Regulate Vacuole Fusion During Tethering and Docking

doi: 10.1101/2020.02.17.953331

Figure Lengend Snippet: Vacuoles isolated from wild-type and elo3 Δ BY4742 yeast were incubated in the absence (A) or presence of [0.5% (w/v) Brij98 (B) or 0.5% (w/v) Lubrol WX (C)] at 4°C and separated through a 0-40% (w/v) Optiprep floatation gradient. Samples collected from the top (fraction 1) to the bottom (fraction 10) of the gradient and subjected to SDS-PAGE and immunoblot analyses for the indicated fusion proteins. (D) Vacuoles isolated from the elo3 Δ yeast are more fluid than wild-type vacuoles. Isolated vacuoles (50 μL of 0.5 mg/mL vacuole isolation) were mixed with 50 μL of 20 μM PDA in PS+pluronic F127. After incubation in the dark for 20 minutes at room temperature, the fluorescence of PDA species (monomer/excimer) was measured (λ ex = 355 nm, λ em (monomer) = 410 nm, λ em (excimer) = 520 nm). The membrane fluidity is then represented as the ration of excimer/monomer fluorescence. Chlorpromazine (CPZ, 10 μM) was included to artificially increase the fluidity of vacuoles isolated from wild type yeast. Error bars represent SEM (n=3). ** p<0.01 (unpaired t test).

Article Snippet: Following isolation, 270 µL of 0.33 mg/mL vacuoles in PS buffer were gently mixed with 30 µL of 10% Triton X-100, 5% Brij98 (ACROS Organics), 5% Lubrol WX (MP Biomedicals), or PS buffer to reach the desired composition (0.5-1%).

Techniques: Isolation, Incubation, SDS Page, Western Blot, Fluorescence