Purification Optimisation - (Jun/04/2008 )
I'm new to this, i need to learn how to optimise protein purification using different conc. of Imidazole (step by step, please). What is the best way to approach this, preferably to start with mini-scale (in 1.5mL eppendorf tube)... does anyone got a protocol that can share with? My protein is in inclusion body. Do i need to have several columns for each immidazole conc ? Please advice.
Check out the protocols in this link:
There's some spin columns from Quiagen to make small purification but yes it's better to run pilot experiments with different conditions like Temerature of culture during expression, Amount of IPTG, Lenght of induction time, type of buffer for extraction (native or denaturing)
Thank you very much for the info. Highly appreciated!
i like this manual (see attachment), learn it wisely