Expermentalists needed for mechanism - (Oct/07/2011 )

Hi,

I have narrowed down two possible mechanisms for my favorite protein. One mechanism involves a charged intermediate (which I can trap) and the other uncharged intermediate. - Does anyone have any idea for experiments to distinghuish between a +1 and neutral - the enzymes size is 35kDa

Thank you.

Would native PAGE be an option? Unlike SDS-PAGE, native separates by size and charge

urea-page should work. urea enhances charge separation. we used to determine the phosphorylation state of a protein with this method.