Figure 2From: Identification and molecular characterisation of a fibrinogen binding protein from Streptococcus iniae.Dot blots showing binding of biotinylated fibrinogen by S. iniae. Strains QMA0076 and QMA0141 were incubated with biotinylated fibrinogen, washed and transferred to PVDF, then membranes were probed with streptavidin-alkaline phosphatase conjugate and developed with NBT/BCIP. Panel A) Lanes 1 and 2 comprised 5 μl cells grown in vegetable peptone, whilst lanes 2 and 3 contained 5 μl cells grown in barramundi serum. Cells were harvested and washed in TBS before being incubated with biotinylated human fibrinogen (Lanes 1 and 3) or TBS (lanes 2 and 4) for 20 min, prior to extensive washing and transfer onto PVDF membrane for subsequent detection. Fb indicates serial two-fold dilutions of biotinylated fibrinogen commencing at 2.5 μg/mL. Panel B) shows serial 2 × dilutions of 5 μl cells diluted in TBS. Lane F, cells incubated first with TBS then with biotinylated fibrinogen before transfer to the membrane. Lane P+F, cells incubated with barramundi plasma followed by biotinylated fibrinogen, Lane TBS, cells incubated twice in TBS.Back to article page