electrophoresis of amino acids

Hi all,

I am much more of a chemist than a biologist and I have been asked to put together an experiment of separation of amino acids using electrophoresis at different pH, does anyone have any methods or advice that I could use.

It would be a massive help as the tutor doesn't know how to do it themselves, I have done the paternity kits so I have a general Idea but not very strong.

Thanks

Ste
 

ajmc98

Don't lick, smell or touch it and you won't die.
Whats his reason for the electrophoresis? just for separation or for zwitterion and isoelectric stuff?
 

ajmc98

Don't lick, smell or touch it and you won't die.
Right, its a bit weird doing it at different pHs for separation but should be the same procedure as the paternity kit, the only thing is depending on the amino acid and the pH it will either form a +ve at low pH or -ve at high pH or neutral at it's isoelectric point and then it varies which way it will run on the gel based on it's charge either towards the anode or cathode so i've seen most loaded from the middle of the gel instead of an edge like with DNA. If you know what amino acids you have you could have a look at their isoelectric points and figure out which ones will go what way.
 
Right, its a bit weird doing it at different pHs for separation but should be the same procedure as the paternity kit, the only thing is depending on the amino acid and the pH it will either form a +ve at low pH or -ve at high pH or neutral at it's isoelectric point and then it varies which way it will run on the gel based on it's charge either towards the anode or cathode so i've seen most loaded from the middle of the gel instead of an edge like with DNA. If you know what amino acids you have you could have a look at their isoelectric points and figure out which ones will go what way.
Thanks that's very useful, do I just prepare the amino acid samples in TAE buffer?
 
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