Phosphoamino acid analysis:Mark Kamps's method磷酸化氨基酸分析【Salk Institute】

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Recipes:

In general, you will need 2 liters of each buffer.

pH 1.9 buffer.

88% Formic acid	50 ml
Acetic acid	156 ml
H2O	1794 ml

Don't use the 98% formic acid and don't adjust the pH.

pH 3.5 buffer

Pyridine	10 ml
Acetic acid	100 ml
100 mmolar EDTA	10 ml
H2O	1880 ml

Don't bother to adjust the pH.

There have been occasional problems with badly smeared PAAs during electrophoresis at pH 3.5. Something seems to leach out of the wicks and make the PAAs relatively insoluble. Tony thinks it is aluminum. Bart thinks that it's calcium. In any case, this problem can be prevented by including 0.5 mM EDTA in the 3.5 buffer.