h******n
发帖数: 493 | 1 This approach is good in theory but whether it will work or not
largely depends on your luck and how well all these experiments
are performed.
I guess you have the sequence of this protein and you have some
idea of which try residues may be phosphorylated.
First thing you want to do is to pufify your protein and then
probe your protein with antibody specific to pY on western blot.
If you see it lights up on the western blot, most likely you
have some pY residues in your protein.
Then you proceed | k********u
发帖数: 69 | 2 actually, it's a quite popular the hot topic right now in MS field. I'm gonna
do similiar stuff. I'm reading the relating papers. As far as I know, there
are even non-gel based method to do this kind of stuff. You ignored how
powerful mass spectrometer can do nowadays. There are tons of ways to
identified phosphoproteomics and one of the simple one is to detech whether
there's a ion that lost 98 m/z or 49 for doubly-charged or 33 for
triply-charged ions. Anyway, read some reviews on phosphoprote | h******n
发帖数: 493 | 3 If you take a pY peptide and try this, you will not be able to
see the loss of phosphate group easily. The phosphate group on
the Tyr residue is really stable. However, this phenomenon is
true for pT and pS peptides because they are much less stable.
Mass Spec is really powerful, but it can't do everything, even
though I am a MS person and I really hope it is that powerful.:)
【在 k********u 的大作中提到】
: actually, it's a quite popular the hot topic right now in MS field. I'm gonna
: do similiar stuff. I'm reading the relating papers. As far as I know, there
: are even non-gel based method to do this kind of stuff. You ignored how
: powerful mass spectrometer can do nowadays. There are tons of ways to
: identified phosphoproteomics and one of the simple one is to detech whether
: there's a ion that lost 98 m/z or 49 for doubly-charged or 33 for
: triply-charged ions. Anyway, read some reviews on phosphoprote
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