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OMIP-007

Phenotypic analysis of human natural killer cells

Micheal A. Eller, Jeffrey R. Currier Cytometry PART A, Volume 81A, Issue 6, 447-449 (2012)

PURPOSE: This panel was developed to characterize the phenotype of human natural killer (NK) cells from cryopreserved peripheral blood mononuclear cells (PBMC) isolated from ACD or EDTA anticoagulated whole blood or apheresis units (Table 1). The application of this panel was to identify changes in NK cell subsets with regard to receptor expression, maturation, homing potential, and activation in the setting of primary HIV-1 natural infection. However, this panel may be applied to a wide variety of disease and normal conditions to characterize NK cells in humans. The performance of this panel was tested on fresh and frozen PBMC as well as using a whole blood lyse no wash procedure.

CELL TYPE: Fresh and Cryopreserved Peripheral Blood Mononuclear Cells

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Aqua Viability
CD3 Human S4.1 PE-Texas Red Exclusion
CD4 Human SFCI12T4D11 ECD Exclusion
CD14 Human Tuk4 PE-Cy5 Exclusion
CD19 Human SJ25-C1 PE-Cy5 Exclusion
CD16 Human 3G8 Pacific Blue NK subset
CD56 Human NCAM16.2 PE-Cy7 NK subset
CD8 Human SK1 APC-H7 NK subset
Integrin alpha 4 beta 7 Human ACT-1 Qdot 655 Homing
CD62L Human DREG56 Qdot 605 Homing
CD158 Human HP-MA4 PerCP-Cy5.5 KIR receptor
CD158b1/b2,j Human DX27 PE KIR receptor
CD158e Human DX9 Alexa Fluor 700 KIR receptor
HLA-DR Human G46-6 FITC Activation
CD57 Human hCD57 APC Differentiation