Donkey anti goat igg conjugated with fitc

Surface expression of TLR2/TLR4 was determined by using differentiated THP-1 cells treated with various concentrations of rRv3131. Cells were harvested after a period of 24 hrs and incubated with goat anti-human TLR2 (2.5 μg/million cells) and goat anti-human TLR4 (R&D, MN, USA) or isotype-matched anti-goat mouse IgG antibodies for 60 min at 4 °C followed by incubation with donkey anti-goat IgG conjugated with FITC (0.5 μg/million cells) (Santa cruz, CA, USA) at 4 °C for another 45 min. Finally, cells were washed and re-suspended in 1× PBS containing 2% FBS. Fluorescence intensity was measured using BD FACS Canto II flow cytometer (BD Biosciences, NJ, USA). Untreated and stained cells were taken as negative control in both the assays. The data were analysed by Flow Jo software (Tree Star Inc., USA).