Blood eosinophils were purified (to >95% purity) from healthy control subjects by using Percoll gradient separation and the CD16 magnetic bead negative selection system (Miltenyi Biotec, Bergisch Gladbach, Germany), as previously described. Written informed consent was obtained from all blood donors, and the study was approved by the Institutional Review Board of Cincinnati Children’s Hospital.
Cd16 magnetic bead negative selection system
Manufactured by Miltenyi Biotec
Sourced in Germany
The CD16 magnetic bead negative selection system is a laboratory tool used to isolate specific cell types from a mixed cell population. The system utilizes magnetic beads coated with antibodies against the CD16 antigen to remove CD16-positive cells, leaving the desired cell population behind.
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2 protocols using cd16 magnetic bead negative selection system
1
Purification of Blood Eosinophils
2
Isolation and Culture of Murine Eosinophils
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Eosinophils were isolated from murine bone marrow of the control group using Percoll density gradient separation and the CD16 magnetic bead negative selection system (Miltenyi Biotec GmbH). Eosinophils were cultured in RPMI 1640 medium (Invitrogen; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Invitrogen; Thermo Fisher Scientific, Inc.) and 1% (v/v) penicillin–streptomycin solution (Gibco; Thermo Fisher Scientific, Inc.) in a humidified atmosphere containing 5% CO2 at 37°C.
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