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"isolation" of haemoglobin

Fred Reyers KPAT1 at OP1.UP.AC.ZA
Thu May 18 08:51:21 EST 1995


Hi Diagnost Users,

I have a small problem in "alchemy"

I am planning a trial where I want to add varying 
concentrations of haemoglobin (hemoglobin for those West of the 
Atlantic) to fluids with predetermined concentrations of various 
chemical constiruents to evaluate what the effect of haemolysis would 
be on a range of analytical methods.

In order to do this I need a fairly pure source of canine haemoglobin.

I can wash erythrocytes (several times), lyse them and this gives me 
a solution of Hb, but it also contains many other erythrocyte 
metabolites.
I can now precipitate the Hb (with TCA or Ammonium Sulphate), filter 
it and NOW comes my problem: How do I reconstitute the precipitated 
Hb (so that it at least resembles the Hb in haemolysed serum samples) 
from a TCA or Amm Sulphate precipitation?
Can anyone out there help???

Thanks in anticipation of a flood of replies,

Fred

Fred Reyers (Prof & Head: Section of Clinical Pathology)
Department of Medicine
Faculty of Veterinary Science
P.Bag X04, ONDERSTEPOORT, 0110,
Rep. of South Africa
Tel: 27-12-529 8288
FAX: 27-12-529 8308 or 27-12-344 2107 (Home)

                       "The great tradgedies of science
            are the slaying of beautiful hypotheses by ugly facts"
                              (Huxley)



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