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PMID: 6815277 Published · ppublish English Journal Article

Cell sorting using a universally applicable affinity chromatography matrix: solid-phase anti-fluorescein isothiocyanate antibody.

Journal of immunological methods ·Vol. 53 ·No. 3 ·1982-09-30 ·Pages 321-34

Baran MM, Allen DM, Russell SR, Scheetz ME, Monthony JF

Abstract

Procedures are described for fractionating cells utilizing a universally applicable cellular affinity chromatography matrix. The affinity matrix consists of immunoabsorption purified goat anti-fluorescein isothiocyanate antibody coupled to large derivatized polyacrylamide beads. This matrix may, in principle, be used to isolate any cell subpopulation provided it has a fluorescein-labeled ligand on its surface. In this report the matrix was used to isolate viable purified fractions of mouse surface Ig-positive cells, Lyt1 cells, and mouse lymphocytes that bind the lectin soybean agglutinin. A preliminary experiment using the anti-FITC beads suggested that this technique can provide a fraction of cells enriched in antigen binding cells. Cell populations isolated by this technique retain their ability to respond to in vitro mitogen stimulation, as well as their ability to be maintained in cell culture following fractionation. Additional experiments using a column consisting of goat anti-rabbit Ig antibody coupled to the same support material are also reported.

MeSH Terms
Animals Antibodies Cell Separation/methods Chromatography, Affinity/methods Female Fluorescein-5-isothiocyanate Fluoresceins/immunology Immunologic Capping Lymphocyte Activation Mice Mice, Inbred ICR Mice, Inbred Strains Receptors, Antigen, B-Cell/isolation & purification Thiocyanates/immunology
Chemicals
Antibodies Fluoresceins Receptors, Antigen, B-Cell Thiocyanates Fluorescein-5-isothiocyanate
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Baran M M
Allen D M
Russell S R
Scheetz M E
Monthony J F
Article Info
Journal
Journal of immunological methods
Abbr.
J Immunol Methods
ISSN
0022-1759
Published
1982-09-30
Pages
321-34
Language
English
Region
Netherlands
NLM ID
1305440
Subset
IM
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