protein aggegation or?
Posted 23 June 2004 - 06:10 AM
We have played with various protein amounts, added detergents ( increasing concentrations of NP40, sarkosyl or SDS), NaCl (150mM), MgCl... Nothing worked except when 0.1% SDS was added which seemed to push p65 slightly in the gel, but too high for the expected size, when compared to native protein marker. Strangely, conditions for purification from E.Coli and insect cells are identical and dyalized/stored in the same buffer...
Is anybody having any sparking idea?
Posted 27 June 2004 - 08:47 AM
Posted 27 June 2004 - 11:55 AM
Please donot discount the possibility of modification though; because NF-kB will be inherently toxic to the insect cell line due to its dynamic biological capabilities and the only way a cell expressing a foreign gene can neutralize the toxicity is by modifying the expressed agent.
Are you sure it is not modified? I am not speaking about the native protein or known research data about NF-kB; just curious about the protein coming off the insect cell line.