| |||||||
| Register | Search | Today's Posts | Mark Forums Read |
| Western Blot Forum Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures. |
 |
| | LinkBack | Thread Tools | Display Modes |
|
#1
09-02-2010, 05:40 PM
| ||||||||||||
| ||||||||||||
 recombinant histagged protein and western blot Hello all, I am expressing histag-proteins and after expression I can easily see a extra band within the correct size of my recombinant protein by SDS-PAGE. However, some of them are negative by western blot (anti-histag). Some are positive (in the same reaction, so the system is working). What can be happening? Is it possible the any folding problems or inclusion bodies might be interfering with the western results? I am using: - For expression: Gateway (invitrogen) with pDEST 17 (C-terminal 6his) and BL-21 cells. - Western: primary Antibody - mouse anti-His (Invitrogen) secondary - ant- mouse-HRP TMB to develop  |
|
| Tags |
| blot , gateway , histag , histagged , protein , recombinant , recombinant proteins , western , western blot |
| Thread Tools | |
| Display Modes | |
|
|
Similar Threads | ||||
| Thread | Thread Starter | Forum | Replies | Last Post |
| Western blot secreted protein control | alphabetty | Western Blot Forum | 4 | 02-20-2011 12:33 PM |
| Human Cytome Project - Update 24 Jan. 2005 | Peter Van Osta | Cell Biology and Cell Culture | 1 | 08-01-2010 02:18 PM |
| Human Cytome Project - an idea - Update 19 April 2005 | Peter Van Osta | Cell Biology and Cell Culture | 1 | 06-01-2009 02:17 PM |
| A Human Cytome Project - an idea - Update 14 March 2005 | Peter Van Osta | Cell Biology and Cell Culture | 0 | 03-14-2005 01:27 PM |
| Human Cytome Project - Update 6 Jan. 2005 | Peter Van Osta | Cell Biology and Cell Culture | 0 | 01-06-2005 10:18 AM |
Linear Mode
