I pulled this out of one my files for the phosphorylated fragments. These
are some of the peptides that I use for standards. I challenge those that
are using IMAC enrichment to successfully bind AND release the last fragment
(with 3 phospho-serines)from their IMAC resin and metal of choice. The fact
that I can't pull out that standard with 3 phosphoserines or successfully
bind a lot of phosphorylated fragments that are devoid of Glutamic acids and
Aspartic Acids makes me uneasy to rely upon this technique exclusively for
finding ALL of the phosphorylated peptides within a digest.
Has anyone had success using any of the (sequence non-specific)
anti-phosphoserine or anti-phosphothreonine antibodies that are out there?
I would really love to hear about your successes or failures. Maybe an
immobilized column of this type would be a better choice to isolate
(digested) phosphorylated fragments.
Peptide Sequence
M+1Ave M+1MI
Delta sleep peptide Nova biochem
#A19312 (S) WAGGDApSGE 929.81 929.30
PKA phospho Ser - AnaSpec # 24506 (S) GRTGRRpSIHDIL
1575.67 1574.79
Phospho Thr peptide - AnaSpec 24514 (T) RRREEEpTEEEAA
1585.52 1584.66
Insulin Receptor Frag. Sigma # P-6462 (Y) TRDIpYETDYYRK
1703.75 1702.75
MARCKS substrate AnaSpec # 10832 (3xS)
3321.76 3319.78
KKKKKRFpSFKKpSFKLSGFpSFKKNKK
I also obtained a list of phospho-tyrosine fragments from
BaChem Americas (www.bachem.com)
M-2010 H-Glu-Asn-Asp-Tyr(PO3H2)-Ile-Asn-Ala-Ser-Leu-OH
H-2706 H-Asn-Pro-Glu-Tyr(PO3H2)-OH
N-1480 Ac-Asp-Tyr(PO3H2)-Val-Pro-Met-Leu-NH2
M-2170 Ac-Ile-Tyr(PO3H2)-Gly-Glu-Phe-NH2
H-3724 Ac-Tyr(PO3H2)-Glu-Glu-Ile-Glu-OH
H-2062 Cyclo(-Gly-Tyr(PO3H2)-Val-Pro-Met-Leu)
H-2532 Ac-Tyr(PO3H2)-Tyr(PO3H2)-Tyr(PO3H2)-Ile-Glu-OH
H-8905 H-Glu-Pro-Gln-Tyr(PO3H2)-Glu-Glu-Ile-Pro-Ile-Tyr-Leu-OH
H-1546
Biotinyl-e-aminocaproyl-Glu-Pro-Gln-Tyr(PO3H2)-Glu-Glu-Ile-Pro-Ile-Tyr-Leu-O
H
H-8900 Biotinyl-e-aminocaproyl-Tyr(PO3H2)-Glu-Glu-Ile-OH
M-2035 H-Thr-Glu-Pro-Glu-Tyr(PO3H2)-Gln-Pro-Gly-Glu-NH2
L-2075 H-Trp-Phe-Tyr-Ser(PO3H2)-Pro-Arg-pNA
Gary Lange
Pharmacia
gary.w.lange@monsanto.com
-----Original Message-----
From: JWU388880@cs.com [mailto:JWU388880@cs.com]
Sent: Wednesday, September 13, 2000 10:25 PM
To: Recipients of ABRF List
Subject: phosphorylated peptides
Does anyone know the commercial sources for phosphorylated peptides (on S or
T)? I appreciate your imput.
Jeff
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