1 reply to this topic

[saad]

Dear all

I've been trying to refold protein and by looking to the literature I found the dilution is a good way for refolding.
I diluted my protein in refolding buffer 40 times then I freeze-dry the sample I want to resusped the protein in different buffers and check if is folded/aggregated etc. Is this the right way to do it beacuse I couldn't find a protocol for refolding protein by dilution.
I know we could use Ion exchange chromatography but we haven't a column suitable to deal with 500-600 ml after dilution, we need high flow rate etc.
Could you help me to find protocols to try refolding a protein by dilution.

Thanks

[protolder]

saad, on 22 March 2011 - 07:24 PM, said:

Dear all

I've been trying to refold protein and by looking to the literature I found the dilution is a good way for refolding.
I diluted my protein in refolding buffer 40 times then I freeze-dry the sample I want to resusped the protein in different buffers and check if is folded/aggregated etc. Is this the right way to do it beacuse I couldn't find a protocol for refolding protein by dilution.
I know we could use Ion exchange chromatography but we haven't a column suitable to deal with 500-600 ml after dilution, we need high flow rate etc.
Could you help me to find protocols to try refolding a protein by dilution.

Thanks

Hola, I can´t give you any protocol wich would be in any book or perhaps in any of the science forums wich all have protocols. You can dilute as you do quickly or dropping diluting buffer that is similar to dialyze, but if you liophylize you will get a high salt concentration. In other time we dialyze against 20mM NH4CO3H at the same pH, wich evaporates in lyophylization. If I found any protocol of refolding by dilution,I´ll send you. Buena suerte