Preparative Electrophoresis of Human Lymphocytes I. Purification of Non-Immunoglobulin-Bearing Lymphocytes by Electrophoretic Levitation

Cf, Gillman; Pe, Bigazzi; Pm, Bronson; Cj, Van Oss

doi:10.1080/00327487408061547

Cited by 11 publications

(7 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the absence of the electric field, these cells sedimented a distance equal to 5% of their electrophoretic migration dis tance (because of the high density range of the gradient) and provided a narrow unimodal distribution profile. It has been suggest ed that certain types of column electropho resis (electrophoretic levitation) in addition to separation by surface potential differences may partly discriminate according to cell size [2]. Our results suggest that in our density gradient electrophoresis system (Fi coll-inverse sucrose density gradient) and for lymphoid cells from humans or animal species, the contribution of sedimentation velocity at unit gravity is minimal.…”

Section: Discussionmentioning

confidence: 59%

“…In the past we applied a newly developed method, namely density gradient electrophoresis [1][2][3][4], for the preparative separation of functionally and phenotypically distinct sub populations of lymphoid cells from animal systems [4][5][6][7][8] and humans [9][10][11]. In this paper, using the same method, we report: (1) studies on the la-positive cells in the mouse spleen, (2) studies on the responding and stimulating cells in mixed lymphocyte culture, (3) the differential mobility of mouse spleen macrophages from T and B lymphocytes.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Density Gradient Electrophoresis of Mouse Spleen Lymphocytes

Platsoucas¹,

Robbins²,

Catsimpoolas³

1980

Int Arch Allergy Immunol

View full text Add to dashboard Cite

Preparative density gradient electrophoresis has been employed for the separation of BALB/c mouse spleen T and B lymphocytes, on the basis of their surface charge. The high mobility cells were found to be predominantly T lymphocytes, whereas the low mobility cells were B lymphocytes. One way mixed lymphocyte cultures were prepared using electrophoretically separated BALB/c lymphocytes, either as responding or stimulating cells and unfractionated CBA/H/T6j mouse spleen lymphocytes, in the appropriate combination. The responding lymphocytes were found only among the high mobility cells which are primarily T lymphocytes. In contrast, the stimulating cells were found in the intermediate and low mobility fractions which contain the B lymphocytes and macrophages. Minimal stimulation in mixed lymphocyte culture was observed by T cells in this allogeneic system. Ia-positive cells were found only in the intermediate and low mobility fractions, which exhibited the capacity to stimulate allogeneic cells in MLC. Macrophages, identified by nonspecific esterase staining, exhibited intermediate electrophoretic mobility, their peak height coinciding with the maximum observed stimulation.

show abstract

Section: Discussionmentioning

confidence: 59%

mentioning

confidence: 99%

Density Gradient Electrophoresis of Mouse Spleen Lymphocytes

Platsoucas¹,

Robbins²,

Catsimpoolas³

1980

Int Arch Allergy Immunol

View full text Add to dashboard Cite

show abstract

“…19 ' 20 Density Gradient Electrophoresis Density gradient electrophoresis has been used for protein and virus separation 21 and has been shown to separate cells on the basis of electrophoretic mobility as measured by microscopic electrophoresis. 30 ' 31 The use of a density gradient counteracts three effects of gravity: 1) particle sedimentation, 2) convection, and 3) zone, or droplet, sedimentation. 24 Downward electrophoresis in a commercially available device utilizing a Ficoll gradient was first used to separate immunological cell types.…”

Section: Role Of Cell Size and Densitymentioning

confidence: 99%

Separation Physics

1990

Low-Gravity Fluid Dynamics and Transport Phenomena

View full text Add to dashboard Cite

Nomenclature A = cross-sectional area of electrophoresis or extraction chamber A M = surface area of separand molecule of particle a -particle radius B = constant of proportionality for partition coefficient b = half thickness of FFE chamber b tj = monomer interaction coefficients C(x) = concentration of density solute in density gradient C f = concentration of the ith solute C fj = osmotic virial coefficient c = concentration of solute D = diffusion coefficient E = electric field strength e = charge on the electron g = acceleration of Earth's gravity Hj = Hjerten number 7 = electric current J t = flow of ith substance K = dissociation constant k = electrical conductivity k,l = coordinates of the sample zone center in horizontal rotation k B -Boltzmann constant L = number of species M = molecular weight m = dimensionless molality N = number of monomers per polymer molecule

show abstract

“…Static column (1)(2)(3) and free flow electrophoresis (FFE) (4.5) are different types of preparative carrier free electrophoresis. They have the common feature that specimens applied remain in fluid medium during fractionation so that particles can freely move, scaling up is possible and material absorption is minimized.…”

Section: Introductionmentioning

confidence: 99%

“…Carrier free electrophoresis is a gentle and powerful method applicable to purify biomaterials on preparative scale and to separate particles such as subcellular organelles or cells (1)(2)(3)(4)(5).…”

Section: Introductionmentioning

confidence: 99%

Interval Carrier Free Electrophoresis for High Resolution Protein Purification

Bauer

Weber

1998

Journal of Dispersion Science and Technology

View full text Add to dashboard Cite

Interval free flow zone electrophoresis is a new mode of free flow zone electrophoresis (FFZE), which facilitates purification of proteins and other molecular substances at very high resolution. It can be performed in the commercially available free flow electrophoresis (FFE) apparatus Octopus. The specimens are loaded and unloaded as usual with the help of a thin buffer film flowing between the two glass plates of the FFE. However, as long as electrical current is applied to a specimen, the medium flow is turned off and conditions of static column electrophoresis are simulated within the FFE device. Thereby electrohydrodynamic flow effects, which widen the sample bands migrating within the electric field, are eliminated while optimal heat removal from the thin buffer film is still possible and a sophisticated technique of harvesting of fractions remains available. Thus interval FFZE offers a gentle preparative method for purification of many kinds of charged molecular species such as proteins or dyes at very high resolution.

show abstract

Preparative Electrophoresis of Human Lymphocytes I. Purification of Non-Immunoglobulin-Bearing Lymphocytes by Electrophoretic Levitation

Cited by 11 publications

References 11 publications

Density Gradient Electrophoresis of Mouse Spleen Lymphocytes

Density Gradient Electrophoresis of Mouse Spleen Lymphocytes

Separation Physics

Interval Carrier Free Electrophoresis for High Resolution Protein Purification

Contact Info

Product

Resources

About