Understanding how the differential binding of Eg5 onto spindle microtubule causes cohesion fatigue during anti-mitotic cancer treatment

Microtubule and kinesins plays multiple important roles in different phases of mitosis. In this study, we aim to characterize the interaction of kinesin-5 to microtubule using Ligand Tracer technology. To do this, we tried to purify kinesin-5 from whole cell lysate by immuno-precipitation. GFP B-2 antibody did not bind to native Eg5-EGFP. After successful immuno-precipitation of HA-Eg5 with HA Y-11 antibody, we encountered difficulties in its elution from the sepharose beads. We managed to immobilize tubulin to polylysine coated glass slides. Ligand tracer experiments were performed using cell lysate containing HA-Eg5 with HA Y-11 FITC as the fluorescent probe. Although there is an increasing signal after addition of fluorescent analyte, the results were not conclusive.Bachelor of Science in Biological Science