I would like to ask if anyone knows if concentrations of Urea 3M and 8M can interfere with protein purification at Ni-NTA and Glutathione Sepharose 43 resins.
Another question, Urea, at 3M and 8M concentrations, may interfere with the efficiency of an indirect ELISA test, where urea would be in the antigen used for sensitization.
High concentrations of urea are compatible with Ni-NTA purification, even at 8M.
I would expect GST affinity purification to be more sensitive to urea because of the possibility of denaturing the GST protein. However, here is a protocol for purification of GST fusion protein from urea-solubilized inclusion bodies, so it can be done.
Article Purification of Proteins Fused to Glutathione S-Transferase
For the ELISA, because of denaturation of the immunoglobulins, my advice would be to keep the urea concentration as low as you can.
I have tested the Urea in Sandwich ELISA, greater than 2M, we got a poor signal.
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