Elemental analysis, Fourier transform infrared (FTIR), 1 H-NMR, fluorescence spectroscopy, and surface tension methods have been used to characterize the molecular structure and the aggregation behaviors of two asphaltenic fractions derived, respectively, from an Algerian petroleum well and a corresponding storage tank deposit. Elemental analysis, FTIR, 1 H-NMR, and fluorescence spectroscopy were used to investigate the chemical composition and structural parameters of asphaltenes, while the surface tension method was used to measure the critical micelle concentration (CMC) in organic solvents with different solubility parameters and polarities in order to characterize the asphaltenes' aggregation behaviors. Results show that the unstable asphaltenes fraction extracted from the storage tank deposit possesses a higher polarity (higher heteroatoms content) and a lower aromaticity than stable asphaltenes from the petroleum well. The CMC results indicate that asphaltenes with high polarity and low aromaticity have a high solubility in polar solvents such as nitrobenzene, whereas asphaltenes with low polarity and high aromaticity are more soluble in solvents with weak polarity, like toluene. It is concluded that the difference of structure of asphaltene samples and polarity of solvents can lead to difference of aggregation behaviors.