Xcell surelock mini cell gel tank

SDS-PAGE was used to confirm the size and purity of the protein. For SDS-PAGE, the protein was loaded onto a NuPAGE™ 4–12% Bis-Tris, 1.0 mm, Mini Protein Gel (ThermoFisher Scientific) and ran on a XCell Surelock Mini-Cell gel tank (Thermo Fisher Scientific) with NuPAGE™ MES SDS Running Buffer (Thermo Fisher Scientific).
Native-PAGE with in-gel fluorescence imaging was used to examine binding between PG65-MIMO and labeled αS. For Native-PAGE, the protein sample was loaded onto a Native-PAGE™ 4–16% Bis-Tris gel and ran on the XCell Surelock Mini-Cell gel tank with Native-PAGE™ Running Buffer (Thermo Fisher Scientific).

A combination of SDS-PAGE and Native-PAGE was used to analyze the aggregation states of the samples. The gels ran on the XCell Surelock Mini-Cell gel tank (Thermo Fisher Scientific), with different running buffers for SDS-PAGE and Native-PAGE.
For SDS-PAGE, total and soluble fractions were prepared and analyzed separately. To prepare the soluble fraction, 50 μL of the sample was centrifuged at 16,900 xg for 5 minutes and the supernatant was used as the soluble fraction. Three μL of Fluorescent Compatible Sample Loading Buffer (Thermo Fisher Scientific) was added to 9 μL of each sample and heated at 95 °C for 15 minutes. Samples were loaded onto a NuPAGE 4-12% Bis-Tris Gel and run at 200 V with NuPAGE MES SDS Running Buffer for 25 minutes (Thermo Fisher Scientific).
For Native-PAGE, 9 μL of the samples were loaded into each well of the Native-PAGE 4-16% Bis-Tris Gel (Thermo Fisher Scientific). Electrophoresis was performed at 150 V with Novex Tris-Glycine Native running buffer for 90 minutes.
Samples containing HiLyte Fluor 488 labeled Aβ42 (green) and Alexa Fluor 647 labeled αS (red), were utilized for fluorescence imaging of SDS- and Native-PAGE gels. The gels were imaged on an Amersham Typhoon RGB (Cytiva, Marlborough, MA, USA) located at the Genomics Core at NYU Center for Genomics and Systems Biology.