AAA - yeast cell lysate

Angela c. Murphy (acmurphy@helix.nih.gov)
Tue, 23 Nov 1999 13:33:42 -0500

I have been asked if I can quantitate a single free amino acid from a
yeast cell lysate. I think I can, with the help of my Beckman 6300,
which uses sodium citrate buffers and post-column ninhydrin detection.
I've asked that lipids be extracted with chloroform and that proteins also
be removed. She's going to try to use ultrafiltration with a low
molecular weight cutoff membrane, about 1,000. What else needs to be
done so that free amino acids are just about the only things left and
things that could mess up my column are removed? What concentrations of
free amino acids are usually seen in yeast cell lysates - nanomoles/ml,
micromoles/ml, millimoles/ml? Excuse my ignorance - I usually work with
peptide and protein hydrolysates, this is my first time with a cell
lysate.
Thanks for any help.
Angela C. Murphy

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Angela C. Murphy, Chemist
Lab. of Cell Biology, NHLBI, NIH
3 Center Drive, MSC 0301, Rm. B1-22
9000 Rockville Pike
Bethesda, MD 20892-0301 USA
tel.: (301) 496-2324
fax: (301) 402-1519
email: acmurphy@helix.nih.gov
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