phosphorilation state detection - (Feb/21/2006 )
Hi,
I would like to know how to detect differences in proteins that may have different grades of phosphorilation. Is it possible by western blot? I heard something about reducing, not reducing conditions. I have 2 mutants with different phosphorilation states ( serines substituted to alanines) How could I differentiate these fhoshorilation states without using radioactivity ? Thanks in advance
-virolo-
QUOTE (virolo @ Feb 21 2006, 12:34 PM)
Hi,
I would like to know how to detect differences in proteins that may have different grades of phosphorilation. Is it possible by western blot? I heard something about reducing, not reducing conditions. I have 2 mutants with different phosphorilation states ( serines substituted to alanines) How could I differentiate these fhoshorilation states without using radioactivity ? Thanks in advance
I would like to know how to detect differences in proteins that may have different grades of phosphorilation. Is it possible by western blot? I heard something about reducing, not reducing conditions. I have 2 mutants with different phosphorilation states ( serines substituted to alanines) How could I differentiate these fhoshorilation states without using radioactivity ? Thanks in advance
Some ideas:
- mass spectrometry
- antobodys for phosphrylated AA
- phosphostain with pro-q diamond phosphoprotein gel stain
- fplc with SCX-columns
Oli
-ms-olli-