SDS Page_Western Blot Notes

Concentration which is revealed in the od 600

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concentration, which is revealed in the OD 600 readings Assuming the cells were inoculated at the same density, the cells density was better for glucose o The GAL1 promoter turns on the one gene expressed in the plasmid o The lightness and darkness of bands just reveals how much protein was extracted from the cell, influenced by the initial concentration of cells used more than anything else o The simply blue stain reveals if we got a good protein extract from the yeast sample, it doesn’t reveal the effect of galactose vs. glucose o In the western blot we did or did not see a band because the protein wasn’t there based on the SDS page results o cut out anything unessential from the figure: met3 mutant transformed with overexpression plasmids induced/uninduced were stained with simply blue Western Blot o In the figure have induced next to uninduced o Take out methods o Put band presence in each lane o In the figure legend include that the primary antibody was for the V5 epitope, V5 is a fusion tag o We saw our protein induced where we expected o
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