GST pulldown- troubleshooting - (Feb/23/2007 )

Hi everybody,

I am doing recently GST pulldown and unfortunately one of my proteins of interest binds unspecific to the beads;
every suggestion will be welcome

thanks

-Tuxy-

I had the same problem as yours, as i posted on the board (Background binding problems in GST pull down). Five of my interested MBP fusion proteins binds to the colum nonspecificly, which are expressed in E. coli in high level, but the other four expressed in relatively low level, do not. I have no idea why. I am still trying with different modification. If you have solution or good idea, I will appreciated if you let me know (zli5@uky.edu). Thanks.

-danxi-

Hi i have got the same problem so if you have the answer, taht would be very welcome for me...camille.foucart@gmail.com.thanks

-camille23-

What is the concentration of salt in the solution where you do the binding?
I had the same problems, but when I used 0.5 M NaCl the binding to the beads disappeared

-rick2000-