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Investigation of the Effects of St. John’s Wort on Cancer and Drosophila HCI Objective 1 : To find out which immune related proteins interact with St. John’s wort or hypericin. 1) Feed 10 drosophila flies with St. John’s Wort for 48 hours. Feed another 10 drosophila flies with their normal food for 48 hours. 2) Grind both sets of 2 sets of drosophila separately in sterile saline. 3) Conduct SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) on the 2 solutions. Compare the 2 gel samples. 4) Extract relevant parts of the gel and send for MALDI-TOF (matrix-assisted laser desorption/ionization time of flight mass spectrometry) @ NUS Objective 2 : To find out whether St. John’s wort affects cancer cell growth 1) Culture Drosophila cancer cell line. 2) Add different concentrations of St. John’s wort (at least 4 concentrations) to media containing cells. Allow incubation. 3) Cell count to check rate of proliferation of Drosophila cancer cells AOS Objective 1: To find out whether St. John’s wort affects cancer cell growth 1) Culture colon cancer cell line. 2) Add different concentrations of St. John’s wort (at least 4 concentrations) to media containing cells. Allow incubation. 3) Cell count to check rate of proliferation of colon cancer cells Objective 2: To find out whether St. John’s wort interacts with norepinephrine, which is found to be responsible for cancer cell growth. 4) Culture colon cancer cell line. 5) Add different concentrations of St. John’s wort (at least 4 concentrations) to media for it to interact with norepinephrine (since norepinephrine is secreted out of the cell). Allow incubation. 6) Add samples from the media into the ELISA, which has wells attached with specific anti-bodies. 7) Allow anti-bodies to bind with the norepinephrine present in the samples. Rinse. 8) Add coloured/fluorescent anti-bodies to bind to the anti-body norepinephrine complexes present in the wells. 9) Measure the fluorescence using the absorbance plate reader. Notes AOS would most probably not be experimenting with hypericin. Liquid St. John’s wort which is water based would be used. (proteins in powdered samples may have denatured during drying)