Pd 1 percp

Manufactured by Miltenyi Biotec

Sourced in Germany

The PD-1-PerCP is a flow cytometry reagent that detects the expression of the Programmed Cell Death 1 (PD-1) protein on the surface of cells. PD-1 is an important immune checkpoint receptor that plays a key role in regulating T-cell activation and function. The PD-1-PerCP reagent is conjugated to the PerCP (Peridinin Chlorophyll Protein Complex) fluorescent dye, allowing for the detection and quantification of PD-1-expressing cells using flow cytometry.

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2 protocols using pd 1 percp

Detailed Lymphocyte Characterization and nAb Quantification

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Heparinized blood was utilized for a 24 h whole blood peptide stimulation assay and for isolation of peripheral blood mononuclear cells using standard density gradient centrifugation protocols. Ethylenediaminetetra-acetic acid whole blood samples were stained and analyzed via flow cytometry (MACSQuant 10, Miltenyi Biotec, Bergisch Gladbach, Germany) to determine lymphocyte counts and immune cell subsets using the following conjugated antibodies: CD8-VioBlue, CD14-VioGreen, CD3-VioGreen, CD3-FITC, CD4-FITC, CD4-PE, CD62L-PE, CD20-PerCP, PD-1-PerCP, CD45RA-PerCPVio770, CD45-APC, and CD56-APCVio770 (Miltenyi Biotec, Bergisch Gladbach, Germany). The percentage of all CD45+ lymphocytes expressing the surface marker of interest were multiplied by the total lymphocyte count to enumerate each lymphocyte subset per unit of whole blood. Serum was isolated from each timepoint and frozen for future analysis to determine SARS-CoV-2 nAb titers using commercially available enzyme-linked immunosorbent assay (ELISA) kits (Cayman Chemical, Ann Arbor, MI, USA).

Smith K.A., Zúñiga T.M., Baker F.L., Batatinha H., Pedlar C.R., Burgess S.C., Gustafson M.P., Katsanis E, & Simpson R.J. (2023). COVID-19 vaccination produces exercise-responsive SARS-CoV-2 specific T-cells regardless of infection history. Journal of Sport and Health Science, 13(1), 99-107.

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Comprehensive Immune Cell Profiling

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Antibodies (CD8 Vioblue, CD3 Viogreen, vγ9 vδ2 FITC, CD4 PE, CD45 Per-CP, vγ9 vδ1 APC, CD56 APC-Vio770, CD16 Vioblue, NKG2D PE, PD-1 Per-CP, CD158b PE-Vio770, NKG2A APC, TCR-γδ APC-Vio770, NKp30 PE) (Miltenyi Biotec, San Diego, CA) were added to 25 µL of whole blood at the proper titrated volumes and allowed to incubate for 30 minutes at room temperature in the dark. Blood samples were lysed with 1X Red Blood Cell Lysis solution (Miltenyi Biotec, San Diego, CA), washed twice in phosphate-buffered saline (PBS), and resuspended in 200 µL PBS.

Mylabathula P.L., Diak D.M., Baker F.L., Niemiro G.M., Markofski M.M., Crucian B.E., Katsanis E, & Simpson R.J. (2022). IL-2 and Zoledronic Acid Therapy Restores the In Vivo Anti-Leukemic Activity of Human Lymphocytes Pre-Exposed to Simulated Microgravity. Frontiers in bioscience (Landmark edition), 27(7).

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