separated two protein
Posted 21 February 2005 - 12:56 AM
Now i can not separate two protein, which showed the closely Mw. Because showed one band on SDS-PAGE, but two band in Native. The native-PAGE, the two band were not so far mobility.
Posted 21 February 2005 - 12:10 PM
I'm not quite sure I understand what your problem is! From what I understand, you try to separate 2 proteins but they are too similar in mass. Is that it?
If so you could do a step of IEF (isoelectrical focusing) prior to SDS-PAGE. 2D-PAGE should get you there.
Posted 21 February 2005 - 08:26 PM
Posted 22 February 2005 - 08:41 PM
Now i try to use Mono Q column for separating two protein, but my enzyme still bound to the column, even used 4 M NaCl in 50 mM Tris-HCl, pH 8.0 + 1 mM CaCl2. If you have some recommentation, please tell me.
Posted 22 February 2005 - 08:54 PM
National Chemical Laboratory,