This chapter provides a comprehensive overview of electrophoretic and chromatographic techniques, emphasizing their pivotal roles in the separation and analysis of complex biological macromolecules in advanced molecular research. Electrophoretic methods, including polyacrylamide gel electrophoresis (PAGE), gradient gels, agarose gel electrophoresis, two-dimensional electrophoresis, isoelectric focusing, and dielectrophoresis, facilitate high-resolution separation based on molecular size, charge, and dielectric properties. These techniques are indispensable for protein profiling, nucleic acid characterization, and the deconvolution of complex biological mixtures. The integration of radiolabeling strategies, involving isotopic tagging and the principles of radioactive decay, significantly enhances the sensitivity of molecular detection. This sensitivity is achieved through rigorous sample preparation and precise quantification via radioisotope counting techniques. Complementing these electrophoretic approaches, chromatographic methods provide high specificity and reproducible separation of intricate biomolecular mixtures. Together, these separation strategies constitute a robust analytical framework essential to molecular biology, proteomics, genomics, and translational biomedical research, advancing both qualitative and quantitative insights into biological systems.