Purification of human basophils by density and size alone
- UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
- Christopher Kepley, Associate Professor (Creator)
- Institution
- The University of North Carolina at Greensboro (UNCG )
- Web Site: http://library.uncg.edu/
Abstract: Basophils typically account for approximately 1% of the white cells in peripheral blood. We have developed a unique method for purifying basophils from whole blood of normal subjects to at least 95% purity. Basophils are separated from other cell types dy density-dependent sedimentation in Percoll and cell sorting, based solely on their size and granularity. The mean overall yield ranged from 5% to 28%. The procedure is typically completed within 4 h. The highly purified basophils obtained are functionally competent and morphologically intact. They release histamine in response to FceRI-mediated stimulation, express FceRI and BSP-1 ligand as analyzed by flow cytometry, and exhibit the known characteristic ultrastructural features of basophils by electron microscopy. This procedure avoids positive-selection antibodies that might perturb receptors on basophils or negative-selection antibodies that might activate other cell types, and can be used to obtain basophils for studies in vitro.
Purification of human basophils by density and size alone
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Additional Information
- Publication
- J. Immunol Methods 1994; 175:1-9.
- Language: English
- Date: 1994
- Keywords
- Basophil, Flow cytometry, Electron microscopy, FceRI, Histamine, Purification