< OMIPS Directory
OMIP-028

Activation panel for Rhesus macaque NK cell subsets

Nicholas Pomplun, Kim L. Weisgrau, David T. Evans, Eva G. Rakasz Cytometry PART A, Volume 87, Issue 10, 890-893 (2015)

PURPOSE: This panel was developed to quantify natural killer (NK) cell subsets in Rhesus macaques (Macaca mulatta) during SIVmac239 infection induced pathogenesis. It includes markers to monitor changes in the activation/proliferation phenotype of up to 12 NK cell populations. The performance of the staining was tested on cryopreserved lymph node samples, and on fresh and cryopreserved peripheral blood mononuclear cells (PBMC) isolated from EDTA anti-coagulated blood. The panel can be used to characterize NK cells in a range of normal and pathologic conditions of this species and can be easily adapted to stain samples from various tissues.

CELL TYPE: Fresh PBMC and lymph node samples

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
Dead cells Macaca mulatta Live/Dead Fix Near IR (775) viability/exclusion
CD3 Macaca mulatta SP34-2 Alexa Fluor 700 exclusion
CD8 Macaca mulatta SK1 Brilliant Violet 510 NK subset
CD14 Macaca mulatta M5E2 Alexa Fluor 700 exclusion
CD16 Macaca mulatta 3G8 Brilliant Violet 711 exclusion
CD20 Macaca mulatta 2H7 Alexa Fluor 700 exclusion
CD45 Macaca mulatta D058-1283 Brilliant Violet 786 hematopoietic cell lineage
CD56 Macaca mulatta B159 FITC NK subset
CD69 Macaca mulatta TP1.55.3 ECD Activation
KLRC1/2 Macaca mulatta Z199 PE-Cy7 NK subset
CD158 Macaca mulatta NKVFS1 PE NK subset
KIR3dl05 Macaca mulatta Brilliant Violet 421 NK subset
Ki-67 Macaca mulatta B56 Alexa Fluor 647 Proliferation
HLA-DR Macaca mulatta L243 Brilliant Violet 650 Activation
PD-1 Macaca mulatta EH12.2H7 Brilliant Violet 605 NK differentiation
NKp46 Macaca mulatta BAB281 PE-Cy5 NK differentiation