This work reports the confluence of static and dynamic fluorescence quenching of rhodamine B (RhB) by chemically exfoliated molybdenum disulfide (MoS2) nanosheets. Both steady state and time-resolved fluorescence quenching measurements were carried out to elucidate the process of energy transfer from RhB to MoS2. The interactive forces investigated through evaluation of thermodynamic parameters from temperature dependent fluorescence measurements are found to be hydrophobic in nature. The negative value of Gibbs free energy (a dagger G) indicates spontaneity of the adsorption process of RhB-MoS2 system. The binding affinity of RhB-MoS2 system is also investigated using UV/Vis spectrophotometer.