Multivalent binding and target crosslinking
Conventional streptavidin can bind multiple biotins and crosslink targets. For membrane-protein motion or clustering studies, distinguish multivalent from engineered monovalent reagents and compare target behavior before and after probe addition.
Interpreting fluorescence intensity
Fluorescence depends on target biotinylation, binding and optical settings. At matched target amount, titrate fluorescent streptavidin and compare positive/negative separation. Report probe amounts and detection settings before interpreting intensity differences as changes in target count.