Anti cd63 pe

Ethanol (>99.8%) was purchased from Merck and type II water was used as an extraction solvent for the solid–liquid extraction process. Thrombin 6 receptor activating peptide (TRAP-6), Adenosine 5′-diphosphate (ADP), and collagen were obtained from Sigma-Aldrich (St. Louis, MO, USA). Sodium citrate vacuum tubes were purchased from Genetics and Technology spa (GENYTEC), Santiago of Chile, Chile. Anti-CD62P-PE, anti-CD61-FITC, and anti-CD63-PE antibodies were purchased from BD Pharmingen (BD Biosciences, San Diego, CA, USA). The HPLC grade solvents used were purchased from Burdick and Jackson (Muskegon, MI, USA).

Stromal vascular fraction (SVF) cells were harvested from the inguinal AT, and to block Fc receptors, they were incubated with FC block (Cat # 156604, TruStain FcX PLUS, anti-mouse CD16/32, BioLegend, London, UK) for 10 minutes. For the macrophages panel, cells were stained with anti-CD86-APC-R700 (Cat # 565479, BD Biosciences, San Jose, USA), anti-F4/80-APC antibody (Cat # 123116, BioLegend), and anti-CD11b PE-Cy7 (Cat # 25-0112-81, eBioscience, San Diego, USA). For the eosinophils panels, anti-Siglec-F BV421 (Cat # 562681, BD Biosciences), anti-CD11b APC (Cat # 101212, BioLegend), anti-CD63 PE(Cat# 564222, BD Biosciences), and anti-CD193 (CCR3) Alexa 647 (Cat# 144508, BioLegend) were used. After staining, cells were washed with a washing buffer (2% FCS in PBS) and analyzed using a BD FACSLyric™ flow cytometer (BD Biosciences).