The immunoprecipitation of B16F10 cell membrane, RBC membrane, membrane mixture, and the fused nano-Ag@erythrosome was conducted using a protein binding immunoprecipitation kit (abs955, Absin) according to the manufacturer’s protocols. The protein amount of all the samples was determined using a BCA kit before Western blot. For each well, the protein amount was quantified to ~40 μg. All the samples were first incubated with anti–Band-3 or immunoglobulin G (IgG)–bound beads (18566-1-AP, Proteintech). The level of gp100 enriched on beads was then measured by Western blotting. Electrophoresis was conducted on 15% SDS–polyacrylamide gel and transferred onto polyvinylidene fluoride membranes (Merck Millipore, USA). Then, the samples were incubated with primary antibody of gp100 (Anti-Melanoma gp100 antibody, ab137078, Abcam), followed by horseradish peroxidase–labeled goat anti-rabbit IgG (H+L) (BA1054, BOSTER). The protein signals were detected with an enhanced chemiluminescence method using an Amersham Imager 600 imaging system (General Electric Company, USA).

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