Dialysis

... at first sight, I hope all the same it's not too stupid to ask.... It can be reduced to "why is it - in this case - necessary to perform a dialysis?" We transfected HEK293T in order to express a his-tagged protein. The cells were kept in DMEM (high glucose, modified with 5% FCS and ...

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... very slowly in several steps. Thus, if you had pH 1 and want pH 8.0, I would do at least eight steps, one pH point per step (i.e. first dialysis against buffer with pH 2, second dialysis with pH 3, third with pH 4...). Does your native protein turn yellow in the reducing buffer. I don't ...

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... of your column? Do you have protein of which you can measure activity? If so, do it and you will see. You will probably have to do some slow dialysis to re-fold your protein.

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... years X = 1,000,000 min = 16,666.67 hrs = 694.44 days = 1.90 years 1. Take some of the E.coli (whole genome is sequenced) and put them into dialysis cup/jar in normal LB broth. 2. Set up three sets of dialysis chambers: a) control – regular LB broth; b) experimental chamber 1: LB broth + ...

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that was just idea, since I know, that proteins are sometimes extracted with the use of urea and then refolded during dialysis. But with the acidic environment it should work as well. Slowly increasing pH and the protein could refold.

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