Considering the clinical importance of Heparin in anticoagulant therapy, intensive efforts have been devoted to develop optical sensor systems for the specific and sensitive detection of Heparin. However, majority of the reported probes either work through turn-off mechanism or are commercially inaccessible. Herein, we report a molecular rotor based dicationic cyanine derivative, commercially known as YOPRO-1, for effective, economic, simple and quick “turn-on” detection of Heparin. For the first time, the formation of emissive H-aggregates of YOPRO-1 has been demonstrated in any chemical or biological environment that offers an excellent sensing platform for the detection of this important biomolecule, Heparin. In addition, YOPRO-1 also undergoes exceptional changes in its absorption features that presents an opportunity of dual read-out i.e., both, colorimetrically aswell as fluorometrically. Apart fromsensitive detection (LOD=0.5nM), YOPRO-1 also shows high selectivity for Heparin as compared to its structurally related analogs. Importantly, the commercial availability of this probe molecule presents an advantage over synthetically challenging probe molecules. More importantly, the dicationic YOPRO-1 shows improved response in high percentage of serum matrix over previously reported monocationic probes and represents a significant advance in the field of Heparin sensing.