Drosophila Raf (D-Raf) is an orthologue of Human B-Raf. D-Raf may have two splice variants, a short (Isoform A, I-A) and a long one (Isoform B, I-B). Protocols for over-expression (E. Coli) and purification of 117 amino acids (aa) N-terminal segment (NTS) of D-Raf protein (I-A), both WT and Mutant (T60E) were established. These protein constructs were also biophysically characterized. Thrombin cleavage conditions were optimized, as monitored by 5 amino acid NTS sequencing, mass spectroscopy and SDS-PAGE electrophoresis. An attempt to purify the 160 aa D-Raf (I-B) NTS failed. NMR characterization in a 700 MHz magnet employing three experiments (HSQC, TOCSY, and NOESY), and circular dichroism spectroscopy indicated that the NTS (I-A) is highly acidic and highly skewed in aa content, but is folded. This establishes a basis for future total assignment and determination of the protein structure.