The separation of cells and organelles by partitioning in two-polymer aqueous phases

Fisher, Derek

doi:10.1042/bj1960001

Cited by 99 publications

(23 citation statements)

References 90 publications

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“…Depending on the choice of phase system, separation can occur on the basis of surface charge, hydrophobic properties or ligand affinity (Albertsson et al, 1982;Fisher, 1981).. This provides an alternative means of organelle isolation to that of other techniques, including centrifugation.…”

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confidence: 99%

Subcellular fractionation of rat liver homogenates using two-polymer phase systems in a toroidal-coil centrifuge

Heywood-Waddington¹,

Sutherland²,

Morris³

et al. 1984

Biochemical Journal

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The principal organelles of rat liver homogenates were fractionated by two-phase partition chromatography using toroidal-coil centrifugation with a mixture of dextran T 500 and poly(ethylene glycol) 6000 in 0.26M-sucrose containing 10mM-sodium phosphate/phosphoric acid buffer, pH 7.4. The effects of varying the following parameters on organelle elution profiles, as reflected by their marker-enzyme activities, were studied: centrifuge speed; the composition and relative proportion of dextran-rich and poly(ethylene glycol)-rich phases in the eluent; flow rate; sample volume; homogenate concentration; helix diameter; tubing bore and the number of loops in the coil. Optimal resolution of the organelles was achieved with a toroidal coil of internal diameter 1.07mm with a 4.55mm helix diameter on a 0.42m-diameter rotor running at 1000 rev./min. The eluent was prepared by combining, in a ratio of 93: 7 (v/v), the poly(ethylene glycol)-rich upper phase and dextran-rich lower phase obtained from a phase mixture containing 3.3% (w/w) dextran and 5.4% (w/w) poly(ethylene glycol). The flow rate of the eluent was 14ml/h. Optimal conditions for separation of the organelles were evaluated. Resolution of plasma membrane and lysosomes was achieved. Separation of endoplasmic reticulum, which showed marked heterogeneity, from plasma membrane was also demonstrated. DNA and marker enzymes for peroxisomes, mitochondria and cytosol showed distinct elution profiles.

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confidence: 99%

Subcellular fractionation of rat liver homogenates using two-polymer phase systems in a toroidal-coil centrifuge

Heywood-Waddington¹,

Sutherland²,

Morris³

et al. 1984

Biochemical Journal

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“…Cell partition is a simple readily avail able method for examining surface proper ties of cells [1][2][3]. It depends on the prop erty of solutions of polyethylene glycol (PEG) and dextran.…”

Section: Introductionmentioning

confidence: 99%

Hydrophobic Affinity Cell Partition Detects Alterations in Surface Properties of Erythrocytes in Rats Bearing a Subcutaneous Leydig Cell Tumour

Gascoine¹,

Dix²,

Fisher³

1983

Pathobiology

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The partition of rat erythrocytes in dextran-polyethylene glycol aqueous two-phase systems containing the hydrophobic affinity probe polyethylene glycol-palmitate has been examined. Erythrocytes from rats bearing the subcutaneous Leydig cell tumour F344 showed a significantly reduced top phase partition compared with erythrocytes from non-tumour bearing control rats, indicating a decreased hydrophobicity of the erythrocyte surface that was associated with tumour growth. Cell partition of readily accessible erythrocytes may offer a simple procedure for following the development of certain tumours.

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“…Partitioning of cells (and membranes) in two-polymer aqueous phase systems depends, in an extremely sensitive manner, on the cells' surface properties and on the phase system's physical properties with which the cells interact [ 1,2]. The physical properties of the phase systems can, to a great extent, be manipulated by the choice of polymer and ionic composition and concentration [1,3].…”

Section: Introductionmentioning

confidence: 99%

Partition behavior of cells and soluble substances in two‐polymer aqueous phase systems

Walter

Anderson

1981

FEBS Letters

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The separation of cells and organelles by partitioning in two-polymer aqueous phases

Cited by 99 publications

References 90 publications

Subcellular fractionation of rat liver homogenates using two-polymer phase systems in a toroidal-coil centrifuge

Subcellular fractionation of rat liver homogenates using two-polymer phase systems in a toroidal-coil centrifuge

Hydrophobic Affinity Cell Partition Detects Alterations in Surface Properties of Erythrocytes in Rats Bearing a Subcutaneous Leydig Cell Tumour

Partition behavior of cells and soluble substances in two‐polymer aqueous phase systems

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