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 a...
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| Format: | Final Year Project (FYP) |
| Language: | English |
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2014
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| Online Access: | http://hdl.handle.net/10356/60306 |
| _version_ | 1826128595784826880 |
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| author | Kwek, Milton Sheng Yi |
| author2 | Li Hoi Yeung |
| author_facet | Li Hoi Yeung Kwek, Milton Sheng Yi |
| author_sort | Kwek, Milton Sheng Yi |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T07:27:30Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/60306 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:27:30Z |
| publishDate | 2014 |
| record_format | dspace |
| spelling | ntu-10356/603062023-02-28T18:04:50Z Understanding how the differential binding of Eg5 onto spindle microtubule causes cohesion fatigue during anti-mitotic cancer treatment Kwek, Milton Sheng Yi Li Hoi Yeung Susana Geifman Shochat School of Biological Sciences DRNTU::Science::Biological sciences 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 Sciences 2014-05-26T07:05:51Z 2014-05-26T07:05:51Z 2014 2014 Final Year Project (FYP) http://hdl.handle.net/10356/60306 en Nanyang Technological University 29 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences Kwek, Milton Sheng Yi Understanding how the differential binding of Eg5 onto spindle microtubule causes cohesion fatigue during anti-mitotic cancer treatment |
| title | Understanding how the differential binding of Eg5 onto spindle microtubule causes cohesion fatigue during anti-mitotic cancer treatment |
| title_full | Understanding how the differential binding of Eg5 onto spindle microtubule causes cohesion fatigue during anti-mitotic cancer treatment |
| title_fullStr | Understanding how the differential binding of Eg5 onto spindle microtubule causes cohesion fatigue during anti-mitotic cancer treatment |
| title_full_unstemmed | Understanding how the differential binding of Eg5 onto spindle microtubule causes cohesion fatigue during anti-mitotic cancer treatment |
| title_short | Understanding how the differential binding of Eg5 onto spindle microtubule causes cohesion fatigue during anti-mitotic cancer treatment |
| title_sort | understanding how the differential binding of eg5 onto spindle microtubule causes cohesion fatigue during anti mitotic cancer treatment |
| topic | DRNTU::Science::Biological sciences |
| url | http://hdl.handle.net/10356/60306 |
| work_keys_str_mv | AT kwekmiltonshengyi understandinghowthedifferentialbindingofeg5ontospindlemicrotubulecausescohesionfatigueduringantimitoticcancertreatment |