Tris glycine protein gels

The adsorption of plasma proteins onto the particle surface was qualitatively characterized using SDS-PAGE. First, sLe^A-coated PLGA, HSA-PLGA, CSPLGA, and HSA-CSPLGA particles were prepared, as described above. Next, 2.5 × 10⁶ sLe^A targeted particles were resuspended in 150 µL of PBS−/− and sonicated at 20% amplitude to disperse. A 500 µL volume of 100% plasma (ACD, HEP, or ACF) was added to the particle solution and incubated for 5 min at 37 °C to mimic flow experiment conditions detailed above. Following incubation, particles were washed with PBS−/− to remove unbound proteins and resuspended in 50 µL of 1X lane marker non-reducing buffer (Thermo Scientific Pierce, Waltham, MA, USA). The particle solution was heated to 95 °C for 5 min using a thermocycler to denature and release surface-bound proteins. The resulting solution was centrifuged at 15,000 rpm for 5 min. Next, 25 µL of supernatant from each sample was pipetted into individual wells on 4–20% Tris-Glycine protein gels (Invitrogen Novex WedgeWell, Waltham, MA, USA). The gel ran for approximately 30 min at 200 V along with a standard molecular weight ladder (Precision Dual Color Protein Standard, Bio-Rad Laboratories, Hercules, CA, USA) for comparison. The gel is removed from the cast and stained with Coomassie blue (Invitrogen SimplyBlue SafeStain) overnight. Gels were washed in deionized water to de-stain and image.