preventing protein degradation

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Thu Sep 25 12:48:31 EST 1997


Yankiwski (yankiwski at aol.com) wrote:
> I'm expressing a protein in Sf9 insect cells using a baculovirus vector. 
> Upon purification of the 6x his-tagged protein on a nickel affinity column
> and after Western analysis, I get multiple bands, with the majority being
> of smaller size and in greater quantity than the full-length protein.

Assuming you don't have protein processing effects there is not much
you can do. Work as quickly as possible in a cold room (check the
temperature :-). In E. coli it also helps to cultivate cells at lower
temperature, but I don't know whether that is possible with Sf9 cells.

--Cornelius.

-- 
/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
/* "Science is the game we play with God to find out what His rules are."  */



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