Picture downloaded from www.bio-rad.comToday we ran a native protein PAGEs to find out the size (molecular weight in Daltons) of our GFP purifications. We used two pre-stained standards: BIO-RAD's Precision Plus Protein Broad Range and Kaleidoscope standards (the specific sizes of the bands in the stantards coming to this blog soon).
Before staining the gel we took pictures using UV light and a fluorescent ruler, so we can measure how much the GFP bands (one from each one of the different amounts of extract, 5, 10, and 15 μl) migrated from the origin.
Once the gels are stained (to make our proteins visible) we will make a new measurement since it is likely the gel will shrink a little bit. Also, other proteins that happen to be hydrophobic as well may show up.
By comparing the migration of our GFP with the migration of the proteins in the standards we will be able to estimate the size of the GFP.
Ideally we should have used more standards and duplicate or triplicate our samples, but for the purposes of this lab what we did is more than enough.
Gels were dried afer staining, following a protocol in which hey are wrapped in cellophane paper
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