| Assay Method Information | |
| | Solid Phase Integrin alphaVbeta6 Binding Assay |
| Description: | Microplates were coated with recombinant human integrin alphavbeta6 (2 ug/mL) in PBS (100 uL/well 25 C., overnight). The coating solution was removed, washed with wash buffer (0.05% Tween 20; 0.5 mM MnCl2; in 1x TBS). The plate was blocked with 200 uL/well of Block Buffer (1% BSA; 5% sucrose; 0.5 mM MnCl2; in 1x TBS) at 37 C. for 2 h. Dilutions of testing compounds and recombinant TGFbeta1 LAP (0.67 ug/mL) in binding buffer (0.05% BSA; 2.5% sucrose; 0.5 mM MnCl2; in 1x TBS) were added. The plate was incubated for 2 hours at 25 C., washed, and incubated for 1 hour with Biotin-Anti-hLAP. Bound antibody was detected by peroxidase-conjugated streptavidin. The IC50 values for testing compounds were calculated by a four-parameter logistic regression. |
| Affinity data for this assay | |
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