antibody production..does it matter if the protein is denatured or not?
Posted 23 June 2009 - 03:06 AM
I was just wondering whether its going to be a problem to send away my induced fragments after theyve been denatured with urea since only then do I remove them from the pellet. Are there any tricks I can use to increase the fragments antigenic properties. Someone suggested its better to send away a gel cut out?
Thank u i advance for any replies!!
Posted 23 June 2009 - 03:36 AM
In my opinion using protein band from SDS-PAGE instead of urea denatured protein makes no sense, since SDS-PAGE is even more denaturing because it's using strong ionic detergent (SDS) and disulfide-breaking reagent(s) (eg. DTT, bME).
You may try to re-nature your protein eg. by rapid dilution. (Three-phase partitioning (TPP) has also some re-naturing properties - contact me for details if you want to try this method).
Posted 23 June 2009 - 07:18 AM
Posted 24 June 2009 - 05:03 AM
Thank u for input