F ab 2 goat anti human igg fc secondary antibody

Relative levels of cell-surface expression of S RBD after transduction of human embryonic kidney (HEK) 293T cells with various S-expressing hAd5 vaccines were determined by flow cytometry. HEK 293T cells (2.5 × 10⁵ cells/well in 24 well plates) were grown in DMEM containing 10% FBS and PSA (100 units/mL penicillin, 100 μg/mL streptomycin, 0.25 μg/mL Amphotericin B) at 37 °C. Cells were either untransduced or transduced with hAd5 S-WT, S-WT + N-ETSD, S-Fusion, or S-Fusion + N-ETSD viral particles at a multiplicity of infection (MOI) of 10. For detection of S RBD 24 hurs after transduction, cells were transferred by gentle pipetting into medium and labeled with an anti-RBD monoclonal antibody (clone D003 Sino Biological) and F(ab’)2-Goat anti-Human IgG-Fc secondary antibody conjugated with R-phycoerythrin (Thermofisher). Labeled cells were acquired using a Thermo-Fisher Attune NxT flow cytometer and analyzed using Flowjo Software.

Parental REH cells and REH cells transduced with CD33^FL or CD33^∆E2 were labeled with CC-96191 or a chimeric version of 9G2 (C2-set domain-directed CD33 mAb [31 (link)]), followed by incubation with an F(ab’)2-goat anti-human IgG Fc secondary antibody (Thermo Fisher Scientific), staining with DAPI, and flow cytometric analysis.