title: Efficient isolation of CD8α positive T cells from postnatal mice using a combined MACS approach.
abstract: Magnetic activated cell sorting (MACS) is a commonly used cell separation technique. Miltenyi Biotec has developed an efficient MACS protocol for isolation of CD8α(+) T cells from adult mice spleen. However, due to a low percentage of CD8α(+) T cells in spleen of postnatal mice, MACS isolation of CD8α(+) T cells results in a low purity. Our study illustrates how this problem can be solved by performing a CD45R(+) B cell depletion prior to a positive selection of CD8α(+) T cells. This protocol can be used when low frequencies of CD8α(+) T cells are present in a tissue also populated with CD45R(+) B cells, and a high purity is required for downstream applications.
authors.affiliation: Institute of Cell Biology and Neurobiology, Center for Anatomy, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Hübner
authors.affiliation: Department of Neurology, Charité - Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Derkow
authors.affiliation: Institute of Cell Biology and Neurobiology, Center for Anatomy, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. Electronic address: anja.braeuer@charite.de.
authors.forename: Anja U
authors.initials: AU
authors.lastname: Bräuer
mesh_terms: D000367:Age Factors; D000818:Animals; D016827:CD8 Antigens / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology*; D018189:Immunomagnetic Separation / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000166:cytology* / Q000276:immunology*
chemical_list: D016827:CD8 Antigens; C431698:CD8alpha antigen
keywords: B cell; Development; Lymphocyte; Spleen