Anti p selectin antibody

Corning HTS Transwell plate system (# CLS3380-1EA) (96 well, with 1 μm pore size polystyrene membrane support) and dimethyl sulfoxide (DMSO) were procured from Sigma (USA). RPMI 1640 medium, penicillin-streptomycin antibiotic (5,000 U Penicillin and 5 mg/ml Streptomycin in 0.9% normal saline), heat-inactivated fetal bovine serum (FBS), EDTA and PBS (phosphate-buffered saline) were purchased from HiMedia (India). Trypsin-EDTA 1× (0.25%) was from Gibco (Ireland). Doxorubicin (Dox) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were the products of Dabur Pharmaceuticals (India) and SRL Chemicals (India), respectively. Anti P-selectin antibody was purchased from BioLegend (USA) and Calcein-AM was purchased from life technologies (USA). Hydroxyurea was the product of Aimcad Biotech (India). All other reagents used were of analytical grade. Type I deionized water (18.2 MΩ. cm, Millipore, USA) has been used throughout the experiments. Flow cyometry was carried out with BD FACSCalibur employing CellQuest Pro software (BD Biosciences, USA). Fluorimetry was performed with multimodal microplate reader (BioTek model Synergy H1, USA). Nanoparticle Tracking Analysis (NTA) was performed on NanoSight LM10 (Malvern, UK).