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. 2023 Jan 10;13:1085892. doi: 10.3389/fphar.2022.1085892

FIGURE 3.

FIGURE 3

Truncated sema3A can interact with CD72 (A). The ELISA plate was coated with 5 μg/mL purified CD72 and increasing concentrations of purified P-sema3A or T-sema3A were added (0-25 μg/mL). Followed by polyclonal goat anti-human semaphorin3A antibody (directed against the N-terminal of sema3A (26-771 amino acids), then by anti-goat horseradish peroxidase antibody, TMB-Plus substrate chromogen, and a stop solution (10% H2SO4). The optical density (O.D.) was measured at 450nm and 360 nm using an ELISA reader and Gen5 software, and the O.D. of each sample was calculated as O.D. sample minus O.D. background (O.D. background = 0.073). The binding curves representing the optical density as a function of P- or T-sema3A concentration were drawn using GraphPad Prism software. (B). The Kd (equilibrium dissociation constant) and Bmax (maximal binding) were analyzed according to one site total binding equation using GraphPad Prism software.