My protein shows 3 bands - Reb1 phosphorylation? (Mar/23/2008 )
Hello,
My first western blot against the yeast Reb1 protein shows 3 discrete bands (close to each other). I wondered if Reb1 has different phosphorylation states that could explain it. If found nothing on the internet. Has someone experienced it also? How can I know what the 3 bands are?
By the way, we're not interested in the phosphorylation states of the protein. I just want to have a nice blot to show in a paper.
Thanks
Max
-maxberge-
QUOTE (maxberge @ Mar 23 2008, 10:08 AM)
Hello,
My first western blot against the yeast Reb1 protein shows 3 discrete bands (close to each other). I wondered if Reb1 has different phosphorylation states that could explain it. If found nothing on the internet. Has someone experienced it also? How can I know what the 3 bands are?
By the way, we're not interested in the phosphorylation states of the protein. I just want to have a nice blot to show in a paper.
Thanks
Max
My first western blot against the yeast Reb1 protein shows 3 discrete bands (close to each other). I wondered if Reb1 has different phosphorylation states that could explain it. If found nothing on the internet. Has someone experienced it also? How can I know what the 3 bands are?
By the way, we're not interested in the phosphorylation states of the protein. I just want to have a nice blot to show in a paper.
Thanks
Max
if you can discriminate your protein in 3 distincts states that will also be a nice blot; you may check bands with anti-phospho-Y or phospho-S/T antibody; if it is really phosphorylation, you may incubate your sample with excessive alkaline or acid phosphatase or Lambda Protein Phosphatase (λ-PPase) to dephosphorylate...
-The Bearer-