I am currently helping a PhD student in his research lab. We have been using Trypsin for the past couple of weeks but I am unsure of its purpose. I was wondering how Trypsin functioned and what it was usually used for? The Trypsin we had been using apparently had "gone bad" and therefore we have to start the experiment over again. What could have caused it to not function properly?
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$\begingroup$ Welcome to Biology S.E.! If you have additional questions, please visit The Help Center $\endgroup$– L.B.Feb 9, 2015 at 23:36
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3$\begingroup$ How do you know that Trypsin has gone bad, when you don't understand it's function in the protocol (no pun intended)? $\endgroup$– Chris ♦Feb 10, 2015 at 6:41
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1$\begingroup$ @Chris You know because the grad student tells you after his cell cultures stop working right. $\endgroup$– user137Feb 10, 2015 at 15:17
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1$\begingroup$ @user137 This can be one of the reasons. $\endgroup$– Chris ♦Feb 10, 2015 at 16:31
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$\begingroup$ I don't see how "bad trypsin" will spoil a cell culture. The only step that trypsin is used in cell culture is for detaching cells from the cell plate/flask, and if the trypsin is not working, the cells will simply not detach, and you will just either add more trypsin or use a different batch of trypsin. $\endgroup$– March HoMar 6, 2015 at 15:12
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Trypsin is a protease, an enzyme that cuts other proteins. It is used in cell culture to help lift cells from a plate by cutting the proteins that hold the cells to the plate and to each other. Trypsin can go bad because a trypsin enzyme can cut other trypsins, eventually destroying the batch's enzyme activity. This is why trypsin must be kept frozen. Of course you have to thaw it out to use it, so it's best to split the trypsin stock into small aliquots, so you don't have to thaw out the whole bottle everytime you split your cells.
