Anti mouse cd105 pe

To measure the mouse CD105 expression, six different cell lines, including HEK293 cells, mEND-HEK293 cells, BNL.CL2 cells, H22 cells, B16 cells, and mTEC cells, were incubated with anti-mouse CD105-PE (eBioscience San Diego, CA, USA) in 1 mL PBS for 30 min, respectively. After incubation, each cell sample was resuspended in PBS for flow cytometric analysis (Beckman/Coulter Epic Elite flow cytometry, USA).
For monitoring the CD105 enrichment, 3 × 10⁵ mEND-HEK293 or HEK293 cells were incubated with selected library pools (250 nM) or aptamer candidates (250 nM) in binding buffer for 30 min at 4℃. To investigate the specificity of the aptamer, 3 × 10⁵ cells of each cell line were incubated with aptamer (250 nM) in binding buffer for 30 min at 4℃. The initial library sequences were used as the negative control. To determine the dissociation constant (K_d) of aptamers, mEND-HEK293 cells (3 × 10⁵) were incubated with aptamer of various concentrations in binding buffer for 30 min at 4℃.
After incubation, each cell sample was resuspended in washing buffer for flow cytometry. The K_d was determined by fluorescence intensity of binding vs the aptamer concentration using the saturation equation Y = B max X / (K_d + X) by SigmaPlot software. The binding assay for each cell sample was repeated three times.