Fig. 5

Binding of purified C. albicans and C. glabrata Tpi1 to human ECM proteins, analyzed using a microplate-based protein-binding assay. Microplate wells were coated with human proteins (3 pmoles per well) in an overnight incubation at 4 ˚C. Solutions of biotin-labeled Tpi1 in 50 μL PBS at a concentration range of 10–800 nM were then added to the walls and the microplates were incubated for 1.5 h at 37 °C. After washing off the unbound protein, the amounts of bound biotinylated Tpi1 were determined using the SA-HRP/TMB system. The values obtained for the control samples, represented by the wells without human proteins, were subtracted from the total binding. Data points represent the mean values of three determinations, with standard deviations