I am seeking to purify a rESAT-6 protein with 6XHis tag.
I purify with Co2+ resin and elute various fractions using imidazole. Because of the future assays I will be performing I am trying to purify this protein without the use of any detergents, which is making this purification a bit challenging.
My rESAT6 is about 12kDa. I am having difficulty finding a SE column that I can use to pull down my protein that is not out of my price range. For example, Superdex pre-packed columns that are running about about $2500 out of my price range. I was wondering if there are any mini-columns out there that I could use to purify this protein.
Any help is appreciated! Thanks!
Hi Jorge
It seems that your proteins are very close together in mass. At which point SEC isn't super useful. I would suggest running an cat or anion exchange depending on the pI of your protein.
Good luck: Emil
You should first improve your separation before attempting ion exchange or size exclusion. Are you running an imidazole gradient? You should try with Zn2+ ions since a lot of contaminant proteins are being adsorbed considerably.
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