Crystal formation requires two steps: nucleation and growth. Nucleation is the initiation step for crystallization. At the nucleation phase, protein molecules in solution come together as aggregates to form a stable solid nucleus. As the nucleus forms, the crystal grows bigger and bigger by molecules attaching to this stable nucleus. The nucleation step is critical for crystal formation since it is the first-order phase transition of samples moving from having a high degree of freedom to obtaining an ordered state (aqueous to solid). For the nucleation step to succeed, the manipulation of crystallization parameters is essential. The approach behind getting a protein to crystallize is to yield a lower solubility of the targeted protein in solution. Once the solubility limit is exceeded and crystals are present, crystallization is accomplished.