Apc cy7 anti human cd3 antibody

Anti-human CD3 mAb (Lymactin-T, 6001T01) was purchased from Cell Science & Technology (Sendai, Japan). Anti-human CD28 mAb (16-0289-85) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Flow cytometry was performed using allophycocyanin (APC)-conjugated IgG₁ (BioLegend), APC–anti-human CD19 antibody (BioLegend), APC–anti human CD107a (BioLegend), APC–Cy7–anti-human CD3 antibody (BioLegend), V500–anti-human CD4 antibody (BD Biosciences, San Jose, CA, USA), and PerCP–Cy5.5–anti-human CD8 antibody (TONBO Biosciences, San Diego, CA, USA).

Flow cytometry was performed with MACS Quant with the following antibodies: human BD Fc Block (BD Biosciences, Franklin Lakes, New Jersey), Viability Dye (eBioscience, San Diego, CA), Pacific Blue anti-human CD14 Antibody (BioLegend, San Diego, CA), FITC anti-human CD19 Antibody (BioLegend, San Diego, CA), PE-Cy7 anti-human CD56 Antibody (BioLegend, San Diego, CA), APC-Cy7 anti-human CD3 Antibody (BioLegend, San Diego, CA), APC anti-human CD16 Antibody (BioLegend, San Diego, CA), BV421 anti-human CD25 Antibody (BioLegend, San Diego, CA), FITC anti-human CD69 Antibody (BioLegend, San Diego, CA), PerCP anti-human CD8a Antibody (BioLegend, San Diego, CA), PE-Cy7 anti-human CD4 Antibody (BioLegend, San Diego, CA), Biotin Rabbit Anti-Mouse FMC63 scFv (BioSwan, Shanghai, China), and BV421-Streptavidin (BioLegend, San Diego, CA).

Antibodies to TLR7, caveolin-1, flotillin-1, MyD88, clathrin, and Ki-67 were purchased from Abcam, Inc. (Cambridge, MA, USA). NFκB antibody was purchased from Cell Signaling Technology, Inc. (Boston, MA, USA). β-actin antibody, HRP-conjugated anti-rabbit, and anti-mouse IgG were purchased from CWBio, Inc. (Beijing, CN). Mouse IgG_2A anti-Human TLR7 PE-conjugated monoclonal antibody and mouse IgG_2A PE-conjugated isotype control were purchased from R&D Systems, Inc. (Minneapolis, MN, USA). BV510 anti-human CD14 antibody and BV421 anti-human CD16 antibody were purchased from BD Biosciences (San Jose, CA, USA). APC/Cy7 anti-human CD3 antibody, PerCP/Cy5.5 anti-human CD8a antibody, FITC anti-human CD56 antibody, PE/Cy7 anti-human CD11c antibody and APC anti-human CD19 antibody were purchased from Biolegend (San Diego, CA, USA). Protease inhibitor cocktails were obtained from Roche Molecular Biochemicals (Mannheim, DE). BCA protein assay kit and PVDF membranes were purchased from Bio-Rad (Hercules, CA). MβCD, media (Dulbecco's modified Eagle's medium, DMEM), MTT and other chemicals were purchased from Sigma (St. Louis, MO, USA). Gardiquimod was purchased from Santa Cruz (California, USA). aPPD was provided by Shanghai Innovative Research Centre of Traditional Chinese Medicine (Shanghai, China). The compound was 97.9% pure as measured by HPLC analysis.