Membrane proteins were obtained from the mitochondrial fraction of HL-60 cells by solubilization with octyl glucoside and bound to heparin-gels. Bound proteins were successively eluted with solutions containing increasing concentrations of Mg2+ in the first and increasing concentrations of Ca2+ in the second chromatography. After SDS-PAGE and subsequent N-terminal amino acid analysis of proteins on each band, 13 proteins were identified. Fifteen out of the 37 proteins analysed were modified at their N-termini. These results show that this two-step affinity chromatography method using divalent cations as eluents can be applied to a variety of membranes for the isolation of specific proteins.
|Number of pages||4|
|Journal||Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences|
|Publication status||Published - 2005 Sept 5|
- Affinity chromatography
- Divalent cations
- Heparin binding proteins
- Membrane proteins