Miltenyi Biotec-tested panel 20 (MBTP 20)
This application protocol describes the flow cytometric analysis of the activation status of NK cells isolated from PBMCs from whole blood of a healthy donor. Immunophenotyping of NK cells was performed on all PBMCs, directly after isolation via CD3 depletion and CD56 enrichment, and after 14 days of culture.
Channel | Specificity | Clone | Purpose | Fluorochrome | Detection filter (nm) (laser) |
V1 | CD56 | REA196 | NK cells | Vio® Bright V423 | 450/50 (violet) |
V2 | CD45 | REA747 | Leukocytes | VioGreen™ | 525/50 (violet) |
B1 | CD335 (NKp46) | REA808 | Activation marker | Vio Bright B515 | 525/50 (blue) |
B2 | CD336 (NKp44) | REA1163 | Activation marker | PE | 585/40 (blue) |
B3 | 7-AAD Staining Solution | - | Viability | - | 655–730 (blue) |
B3 | CD3 | REA613 | Cell type exclusion | PerCP-Vio 700 | 655–730 (blue) |
CD14 | REA599 | Cell type exclusion | |||
B4 | CD337 (NKp30) | REA823 | Activation marker | PE-Vio 770 | 750LP (blue) |
R1 | CD314 (NKG2D) | REA797 | Activation marker | APC | 655–730 (red) |
R2 | CD16 | REA423 | Maturation marker | APC-Vio 770 | 750LP (red) |
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