Hi, Chris!
I have several alternative suggestions:
1) Run cytochrome C with a salt gradient by cation-exchange (@ J. Chromatogr.
266 (1983) 23-37). A fresh sample gives a single peak. As the sample is
exposed to light over a few hours, there is an accumulation of the
photoreduced form that elutes in an earlier peak. You can convert all of the
protein to the reduced form with sodium dithionite or reoxidize it with
potassium ferricyanide.
2) Draw a purple-capped tube of someone's blood and prepare a hemolyzate
(details supplied on request). Resolve the hemoglobin variants by
cation-exchange (@ Clin. Chem. 39 (1993) 820). This will afford a good
quantitation of the Hb A1c peak (relevant to diabetes diagnosis and
monitoring) and Hb A2 peak (relevant to thalassemia diagnosis) as well as the
main Hb A peak. If anyone in the class is a carrier for one of the more
common hemoglobin variants, such as S or C, it will show that too.
Best regards,
Andy Alpert
PolyLC Inc.
9151 Rumsey Road, ste. 180
Columbia, MD 21045
tel: (410) 992-5400 FAX: (410) 730-8340
*************************************************************
<< Subj: laboratory exercise in hplc
Date: 01/10/2000 5:35:49 PM Eastern Standard Time
From: halkidesc@uncwil.edu (chris halkides)
Sender: abrf-request@aecom.yu.edu (Association of Biomolecular Resource
Facilities)
To: abrf@aecom.yu.edu (Recipients of ABRF List)
Hello Everyone,
I am interested in developing a senior undergraduate lab exercise
in the area of the hplc of proteins. Does anyone have any suggestions for
a protein mixture to be be separated? literature references? I would say
that the constraints are commercial availability of proteins and use of
not-too-esoteric columns and reagents. I would think maybe a mixture of
isozymes would be good (perhaps someone has a particular suggestion.
Thanks to all for any help.
Chris
Christopher Halkides
Dept. of Chemistry, UNCW
601 S. College Road
Wilmington, NC 28403-3297
(910) 962-7427
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