precipitation of phospho-protein - (Apr/03/2007 )
Dear all,
I am in trouble: i need to evaluate the phosphorilation of STAT-3 in my cells, so I am preparing to do wb for phospho-stat3. However, due to unexperience in protein handling, I ended up with a low protein concentration (around 300ug/ml). I do have enough protein for the analysis, but i need to concentrate it in order to run a gel. I was wondering if common precipitation methods, such as ethanol or TCA/ethanol, could affect the phosphorilation of my protein.
Can anybody help?
thank you
margherita
-baggins-
QUOTE (baggins @ Apr 3 2007, 10:55 AM)
Dear all,
I am in trouble: i need to evaluate the phosphorilation of STAT-3 in my cells, so I am preparing to do wb for phospho-stat3. However, due to unexperience in protein handling, I ended up with a low protein concentration (around 300ug/ml). I do have enough protein for the analysis, but i need to concentrate it in order to run a gel. I was wondering if common precipitation methods, such as ethanol or TCA/ethanol, could affect the phosphorilation of my protein.
Can anybody help?
thank you
margherita
I am in trouble: i need to evaluate the phosphorilation of STAT-3 in my cells, so I am preparing to do wb for phospho-stat3. However, due to unexperience in protein handling, I ended up with a low protein concentration (around 300ug/ml). I do have enough protein for the analysis, but i need to concentrate it in order to run a gel. I was wondering if common precipitation methods, such as ethanol or TCA/ethanol, could affect the phosphorilation of my protein.
Can anybody help?
thank you
margherita
if you take methanol/chloroform precipition (Wessel & Flügge) should not affect phospho status
-The Bearer-