Hi to all,
If a protein is encapsulated in a liposome, does it respond to colorimetric methods such the Bradford assay? Or if I perform an electrophoresis, does the protein respond to silver staining or coomassie blue?
Thank you.
Antonio,
As far as staining after electrophoresis, the protein should be fine if you are running SDS/PAGE. The denaturation step should disrupt the liposome and coat the protein with SDS, and allow it to run at the proper molecular weight. It will be no different than doing it from a cell lysate. IF you try to run a non-denaturing gel, the liposome may never enter the gel, it may be too large for the pores of the gel or have the wrong exterior charge, depending on the lipids. As to Bradford assays, I am not sure how the lipids may react, or if they will remain as a liposome. They might be disrupted and you would only have to worry about a chemical reaction with the stain interfering with results, or they may remain intact, blocking the reaction. When I do the assay, the package insert had data on the amounts of many materials that can be added without interfering with the assay, that may help.
Does de liposome itself could be stained with silver or Coomassie?
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