Reactions were boiled with the SDS-PAGE loading buffer, loaded onto a 15% SDS-PAGE gel, transferred to PVDF, and probed using a mouse-derived monoclonal anti-polyQ primary antibody and IRDye 680RD Goat anti-mouse secondary antibody for LI-COR image acquisition. Proposed oligomerization states ...
PD is a common chronic neurodegenerative disorder that affects approximately 1% of the population over the age of 65. The clinical manifestations of PD include muscular rigidity, static tremor, bradykinesia, and gait difficulty [1,2]. PD is an age-related disorder of sporadic or familial origin,...
Clones were analyzed by SDS-PAGE and Sanger sequencing [23]. MAGT1-HA-pCI-neo vector was described in Ref. [24]. 2.2. Hypoxic chamber Cells were transferred to hypoxic chamber (Ruskinn Invivo2 400, UK) at 37°C and maintained in deoxygenated culture medium at the following gas ...
Purity of the preparations was >95% as confirmed by SDS-PAGE and MALDI-TOF/TOF. Labeling of the different cysteine αS variants by maleimide chemistry with either AlexaFluor488-maleimide (AF488, ThermoFisher Scientific, Waltham, MA, USA) or TEMPOL-maleimide (Toronto Research Chemicals, Toronto,...
000 ×gat 22 °C for 30 min. Protein concentrations of both fractions were determined by the bicinchoninic acid (BCA) assay (Thermo Fisher Scientific, #23225). Lysates were spiked with 5 × Laemmli containing 0.3 M Tris/HCl pH 6.8, 50% glycerol, 1% SDS, 0.05% bromophenol blue and ...
Cancers 2021, 13, 1974 15 of 19 After the incubation time, the cells were washed three times with ice-cold PBS. Finally, cells were lysed by adding 1% SDS/0.1 M NaOH and shaking for 30 min at room temperature. To quantify the radioactivity in the cell extracts, an automatic gamma ...