Special order research product lsr 2 analytical flow cytometer

Counts of the CD16A-pos NK cells within the PBMCS were as previously described [17 (link)]. Fifty-microliter aliquots of PBMCs were added to TruCount^® tubes (Becton Dickenson no. 340334 [47 (link)]) with fluorescent beads, labeled for 30 min with antibodies, fixed, and analyzed without washing to count beads and cells. The cells were labeled with PacBlue anti-CD45 (clone HI30) to identify all cells; FITC-anti-CD3e (clone UCHT1) to identify T cells; FITC-anti-CD7 (clone CD7-6B7), APC-Cy7 anti-CD56 (clone HCD56), and PerCP-anti-CD16A (clone 3G8) to identify ADCC effector cells, and PE-Cy7 anti-CD33 (clone P67.6) to identify monocytes, purchased from BioLegend (San Diego, CA, USA). The mAbs were all mouse IgG1 that do not bind to human CD16A. The ADCC effector NK cells were distinguished as CD3negCD7posCD16posCD33negCD45pos and CD56variable. Inclusion of anti-CD7 was critical to distinguishing the subgroup of CD56negCD7posCD16Apos NK cells [48 (link)] from CD56negCD7negCD16Apos monocytes (that are largely CD33pos [49 (link)]). Cells were analyzed the same day as the ADCC assays using a BD Biosciences Special Order Research Product LSR II analytical flow cytometer with a high throughput sampler unit. The data were assessed with FlowJo software (FlowJo, LLC, Ashland, OR, USA).