Purification of intact plant protoplasts by flotation at 1g. (49/214)

From a standard plant tissue digest adjusted to a density of 1.07 g/ml, protoplasts can be harvested by flotation through a low density barrier (1.03 g/ml). The delicate nature of these bodies is suited to this flotation strategy which can be carried out at 1g.  (+info)

Purification of peroxisomes using a density barrier in a swinging-bucket rotor. (50/214)

In iodixanol, peroxisomes are the densest organelle in the light mitochondrial fraction and are therefore easily separated from the other components (lysosomes, mitochondria, etc.) in a preformed isosmotic continuous gradient. Because of the large difference in density between peroxisomes and the next densest organelle (mitochondria), a density barrier is effective. The resolution of the peroxisomes is far superior than that in sucrose and, unlike in Percoll, there is no contamination from endoplasmic reticulum.  (+info)

Fractionation of Golgi, endoplasmic reticulum, and plasma membrane from cultured cells in a preformed continuous iodixanol gradient. (51/214)

A continuous iodixanol gradient within the range 0-30% (w/v) iodixanol can resolve the major membrane compartments of the endoplasmic reticulum, Golgi membranes, and plasma membrane from a postnuclear supernatant prepared from a cultured cell homogenate. The precise density range of the gradient and the centrifugation conditions (100,000-200,000 g for 2-16 h) vary with the type of cell and the requirements of the separation. The strategy is widely used to study the processing of proteins within cells.  (+info)

Purification of peroxisomes in a self-generated gradient. (52/214)

In iodixanol, peroxisomes are the densest organelle in the light mitochondrial fraction and are therefore easily separated from the other components (lysosomes, mitochondria, etc.) in a self-generated gradient. Self-generated gradients make sample handling very easy and are highly reproducible but need to be formed in either a vertical, near-vertical, or small volume high-performance fixed-angle rotor. The resolution of the peroxisomes is far superior than that in sucrose and, unlike in Percoll there is no contamination from endoplasmic reticulum.  (+info)

Purification of peroxisomes in a preformed iodixanol gradient in a fixed-angle rotor. (53/214)

In iodixanol, peroxisomes are the densest organelle in the light mitochondrial fraction and are therefore easily separated from the other components (lysosomes, mitochondria, etc.) in a preformed isosmotic continuous gradient. The resolution of the peroxisomes is far superior than that in sucrose and, unlike in Percoll there is no contamination from endoplasmic reticulum.  (+info)

Isolation of a mouse motoneuron-enriched fraction from mouse spinal cord on a density barrier. (54/214)

After a combined enzymic and mechanical disruption of the spinal cord tissue, the low-density motoneurons band at the interface of a 1.06-g/ml barrier through which other contaminating cells sediment.  (+info)

OptiPrep density gradient solutions for macromolecules and macromolecular complexes. (55/214)

Any density gradient for the isolation of mammalian cells should ideally only expose the sedimenting particles to an increasing concentration of the gradient solute. Thus they will experience only an increasing density and viscosity, other parameters such as osmolality, pH, ionic strength and the concentration of important additives (such as EDTA or divalent cations) should remain as close to constant as possible. This Protocol Article describes the strategies for the dilution of OptiPrep in order to prepare such solutions for mammalian cells.  (+info)

Rapid purification of nuclei from animal and plant tissues and cultured cells. (56/214)

Nuclei are isolated by buoyant density banding in a discontinuous iodixanol gradient, under isoosmotic conditions. The low viscosity of the gradient allows the purification to be carried out at 10,000g in only 20 min. The method avoids possible damage to nucleoprotein complexes caused by hyperosmotic sucrose gradients. Although developed for mammalian liver the method can be applied (with or without minor modifications) to any tissue or cell type.  (+info)