Mouse cytokine antibody array c2 kits

Mouse cytokine antibody array C2 kits were purchased from RayBiotech (Norcross, GA, U.S.). The cytokine layout is listed in Supplementary Figure 2. Briefly, cell supernatants from TAMs or mice plasma were collected before and after BM treatment. After antibody array membranes were blocked in 5% BSA for 30 min at room temperature, cell supernatants were cultured with antibody arrays overnight at 4 °C and washed for three times. Biotinylated antibody cocktail was then incubated with the membranes for 2 h, followed by signal amplification with HRP-streptavidin. Finally, the signals were detected by chemiluminescence method with ECL kit purchased from GE Healthcare (Buckinghamshire, U.K.).

Mouse cytokine antibody array C2 kits were purchased from RayBiotech (Norcross, GA). Briefly, TAMs supernatants from TAMs were collected. The antibody array membranes were blocked in 5% BSA for 30 min at room temperature, cell supernatants were then cultured with antibody arrays overnight at 4 °C and washed for three times. Biotinylated antibody cocktail was then incubated with the membranes for 2 h, followed by signal amplification with horseradish peroxidase -streptavidin. Finally, the signals were detected by chemiluminescence method with ECL Advance reagent and quantified using ImageLab software. A triplicate independent experiment was performed.