were coated with recombinant human Nectin-1 extracellular domain (R&D Systems), recombinant human Nectin-4 extracellular domain (R&D Systems), mouse laminin (Invitrogen™), or bovine serum albumin (BSA; Probumin Diagnostic Grade, MilliPore) diluted in PBS. Covered plates were incubated overnight ...
Proteins were eluted using NPI-250 buffer (250 mM imidizole, 300 mM NaCl, 50 mM NaH2PO4 pH 8.0) buffer. Protein concentration was quantified using the Advanced Protein Reagent (Cytoskeleton Inc.), and SDS-PAGE analysis. Screening for Y145X mutations. Random mutations into position Y145 ...