Investigations into the properties of haemoglobin often require the isolation of the valence intermediates (αo)2β+)2 and (α+ βo2)2. Chromatofocusing with an anion-exchange gel (Mono PTM; Pharmacia, particle size 10 μm) in an HR5/20 column at various temperatures (10–25°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, (αo2β+)2, (α+ βo2)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.

Bolzacchini, E., Fermo, I., Rovida, E., Colombo, R., Samaja, M. (1987). Separation of the valence intermediates of human-hemoglobin by high-performance chromatofocusing. JOURNAL OF CHROMATOGRAPHY A, 397(C), 233-237 [10.1016/S0021-9673(01)85006-8].

Separation of the valence intermediates of human-hemoglobin by high-performance chromatofocusing

BOLZACCHINI, EZIO;
1987

Abstract

Investigations into the properties of haemoglobin often require the isolation of the valence intermediates (αo)2β+)2 and (α+ βo2)2. Chromatofocusing with an anion-exchange gel (Mono PTM; Pharmacia, particle size 10 μm) in an HR5/20 column at various temperatures (10–25°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, (αo2β+)2, (α+ βo2)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.
Articolo in rivista - Articolo scientifico
high-performance chromatofocusing; human-hemoglobin; Separation
English
1987
397
C
233
237
none
Bolzacchini, E., Fermo, I., Rovida, E., Colombo, R., Samaja, M. (1987). Separation of the valence intermediates of human-hemoglobin by high-performance chromatofocusing. JOURNAL OF CHROMATOGRAPHY A, 397(C), 233-237 [10.1016/S0021-9673(01)85006-8].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/33687
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