FYI, the hemoglobin from hell has finally been denatured!! Thanks to
everyone who gave me ideas. Turns out that pH IS one of the problems. I
changed the initial pH of 6.8 by adding TFA(final= 0.2%) to one sample,
and increased the pH to 8.8 for another.
I also kept the pH at 6.8 for a third sample, but used the 20mM DTT
instead of 2-Mercaptoethanol. All three worked! How special.
Thanks again,
Karla Somerville-Armstrong
UMBC Department of Chemistry
1000 Hilltop Circle
Baltimore, MD 21250
ksomer1@gl.umbc.edu