Fluram labeling of high density NH2 surfaces

Some surface bonded and fluram labeled primary amines show a new red-shifted emission in their fluorescence spectrum. We compared the fluorescence of fluram bonded to various amine functionalized surfaces including tetraethylene pentamine (TEPA), 1,2-ethylene diamine (EDA), 3-aminopropyltriethoxy silane (APTES) and trimetylol-propanetris-(beta-aziridino)-propionate (ATA). All reactions were also monitored using XPS. It was found that the new spectral features seem to be related to the local density of amino groups on the surface. In order to verify this hypothesis we synthesized a surface bonded dendrimer with a step-wise increasing NH₂ density. Using molecules with different lengths as branches in the dendrimer, the amine density can be varied. Only in the case of the highest density, the new fluorescence emission was detected. Consequently, the fluorescence of fluram coupled to amines at a surface can be used to quantify the concentration of these amines and the appearance of the red-shifted emission indicates a high local density of the amino groups.