β Scribed by Ezio Bolzacchini; Isabella Fermo; Ermanna Rovida; Roberto Colombo; Michele Samaja
No coin nor oath required. For personal study only.
Investigations into the properties of haemoglobin often require the isolation of the valence intermediates (alpha o2 beta+)2 and (alpha+ beta o2)2. Chromatofocusing with an anion-exchange gel (Mono PTM; Pharmacia, particle size 10 micron) in an HR5/20 column at various temperatures (10-25 degrees C) provides an excellent method for this task. A linearly decreasing pH gradient (8 to 7, generated by Polybuffer 96, Pharmacia) eluted sequentially the species methaemoglobin, (alpha o2 beta+)2, (alpha+ beta o2)2 and oxygenated haemoglobin. Calibration graphs help in quantitative analyses. This method is simpler and less time consuming and provides a similar or even better resolution than the traditional ion-exchange or isoelectric focusing methods.
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## Abstract Rare abnormal haemoglobin (Hb) often poses a problem in the diagnosis of relative electrophoresis mobility. The rare neutral Hbs interact with the Hb S and thus can increase the severity of sickle cell anaemia. In the present study, we investigated the use of reversedβphase highβperform