This is probably a dumb question, but what is the purpose for adding an n terminal secretion peptide to my protein insert during transfection into mammalian cells? Why would I want the protein secreted into the media?
Brett Seese If you want to stimulate other cells in your flask because they have an receptor for your protein or because you want to isolate and purify the protein from the supernatants.
Dear Brett Seese
If the protein is secreted on the media, you can avoid cell lysis and protein purification is much more simple because if it is true that you have to load in the coloumn an high volume of culture surnatant, if it is also true that culture surnatants contain very few impurities and you can recover a protein with high purification level with just one step of affinity chromatography which is almost impossible if you are performing cell lisate and purification from intracellular extract and moreover it avoid intracellular accumulation of high level of protein that may be toxic for the cells and limit cell growth and protein expression.
good luck
Manuele
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