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> <<PepSyn:solubilizing and purifying phospho peps>>
attached mail follows:
Hello ABRF peptide guru's!!
I have three peptides that have two phospho Y's each.
One has overall -5 charge(19mer), -3 (19mer), and -2 charge (23mer).
All precipitated during cleavage in their TFA. I have been able to
get the one with -3 charge in solution at about 0.5 mg/mL slightly
acidic water. The others are more like 0.1 and I have 0.25mMol Synth
to purify.
The HPLC traces look like a forest but the Mass specs are pretty
clean. I got the -3charge purified on a c18 but when I run
analyticals of the purified material, it reverts to multiple peaks
again. I checked by MS of the various fractions I got from the prep
and low and behold they have the same mass so I have multiple charge
species that run at different retention times.
I've thought about dissolving the other two in Guanidinium HCl but I
am worried about them ppt'ing during purification on my column let
alone the fact that I will probably get multiple species again
although their analyticals look much better.
Any purification suggestions would be appreciated.
Thanks,
Lisa
Lisa Bibbs, Ph.D.
10550 N. Torrey Pines Rd.
SP3
La Jolla, CA 92039
This archive was generated by hypermail 2b29 : Fri Sep 01 2000 - 16:10:38 EDT