Alexa fluor 488 anti cd56

Reduction in CD16⁻CD56^bright NK cells has been suggested as one of the mechanisms of decreased NKA.19 (link) Thus, we assessed CD16⁺CD56^dim/CD16⁻CD56^bright NK cell subset ratio for some subjects of this study to investigate whether altered NKA is attributable to change of this ratio. The first 20 subjects from each group were selected and their NK cell subset distributions were analyzed using flow cytometry. A 5-mL sample of whole blood was collected in a heparin tube and diluted with PBS in a 1:1 ratio. PBMCs were isolated by centrifugation from the diluted whole blood and overlaid on Histopaque^®1077 (Sigma-Aldrich, Darmstadt, Germany) in a conical tube. These collected PBMCs were washed and resuspended in staining buffer. PBMCs were stained with Alexa Fluor488-anti-CD56, APC-anti-CD16, and PE-anti-CD3 fluorochrome-conjugated monoclonal antibodies (BD Biosciences, Seoul, Korea). After the reaction was completed, the mixture was transferred to a conical tube and analyzed by BD FACSCalibur™ (BD Biosciences). CD16⁺CD56^dim and CD16⁻CD56^bright NK cells were gated from CD3⁻CD56⁺ cell population and the ratio of CD16⁺CD56^dim/CD16⁻CD56^bright was determined.