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An Optimized Immunomagnetic Bead-based Negative Selection Protocol for CD4 T-cell Isolation from Mouse Lymph Nodes and Spleen.
- Source :
- Scandinavian Journal of Immunology; Mar2008, Vol. 67 Issue 3, p285-294, 10p, 2 Charts, 6 Graphs
- Publication Year :
- 2008
-
Abstract
- This study describes an immunomagnetic bead-based methodology for optimal purification of mouse CD4<superscript>+</superscript> T cells. It reproducibly yields highly pure CD4<superscript>+</superscript> T cells from mouse lymph nodes (95–99%) and spleen (93–96%) with no residual antigen-presenting cell (APC) function in the purified population. The recovery of the starting CD4<superscript>+</superscript> T-cell population is consistently high (>70%) and many samples can be simultaneously processed in a short period of time. The key factors responsible for improved purity are combinations of monoclonal antibodies that were found, through trial and error, to yield T cells of maximal purity achievable by non-flow sort-based negative selection. These cocktails efficiently target unwanted cell subsets with antibodies against multiple surface markers expressed by non-CD4<superscript>+</superscript> T cells. Because immunomagnetic bead-based protocols do not require the expensive and cumbersome processes required by flow sort-based purification, the methodology described here should find widespread use. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 03009475
- Volume :
- 67
- Issue :
- 3
- Database :
- Complementary Index
- Journal :
- Scandinavian Journal of Immunology
- Publication Type :
- Academic Journal
- Accession number :
- 28785253
- Full Text :
- https://doi.org/10.1111/j.1365-3083.2007.02066.x