I use gfp agarose from Chromotek to elute the protein while i can only get a denaturing protein. How to elute the protein from beads under native condition since our target protein is an enzyme whose activity need to be maintained to do the further experiment. Thanks a lot, if you can help me solve this problem.
The recomended protocol by Chromotek is to elute using Glycine at pH 2.5 and then neutralise with 1 M Tris pH 10.4. You can then buffer exchange into your chosen assay buffer.
https://www.ptglab.com/products/pictures/pdf/Application_Note_Elution_from_GFP-Trap.pdf
Another way you could use is to add in a protease cleavage site (TEV, HR3C etc) between the GFP and your protein of interest, and then do an on bead cleavage overnight with the protease to release your protein with the GFP remaining bound to the resin. The protease can then be removed by passing the elute over affinity resin against whatever the protease is tagged with (His6, GST etc).
Other than those two options, there's not really much else that you can do. If you want to maintain native conditons, you'll have to switch to an alternate purification strategy with a tag such as His6, GST, strep II etc.
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