We recommend applying the counterstaining protocol after the completion of the Amplification step in section 7.3, step 5 of the Duolink In Situ Fluorescence User Manual. The following protocol exemplifies counterstaining using a directly labeled FITC anti-alpha tubulin antibody:
Perform all the steps at room temperature.
Above:Single recognition of HER2 in SK-BR-3 cells. Red: PLA signals, each representing one HER2 protein. Green: counterstain of alpha tubulin. Blue: nucleus. PLA probes anti-Rabbit PLUS and MINUS, Duolink In Situ Detection Reagents Orange.
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