PBMC Surface Staining for Flow Cytometry

We performed surface staining of freshly frozen PBMC from CTOT-09 study as previously published.^{25 (link)} Anti-human antibodies included: FITC-anti-CD127 (clone: A019D5; Biolegend), PE-anti-CD3 (clone: UCHT1; BD Bioscience), PerCP-Cy5.5-anti-CD57 (clone: HNK-1; Biolegend), PE-Cy7-anti-CD4 (clone: RPA-T4; eBioscience), APC-anti-CD25 (clone: 4E3; Miltenyi Biotec), APC-Cy7-anti-PD-1 (clone EH12.2H7; Biolegend), and BV510-anti-CD8 (clone: RPA-T8; BD Biosciences). Data were acquired (> 1 × 10⁶ events) on a 3-laser FACSLyric flow cytometer (BD Biosciences) and analyzed using FlowJo software.

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Cravedi P., Fribourg M., Zhang W., Yi Z., Zaslavsky E., Nudelman G., Anderson L., Hartzell S., Brouard S, & Heeger P.S. (2020). Distinct peripheral blood molecular signature emerges with successful tacrolimus withdraw in kidney transplant recipients. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 20(12), 3477-3485.

Publication 2020

PE-anti-CD3 PerCP-Cy5.5-anti-CD57 PE-Cy7-anti-CD4 BV510-anti-CD8

Anti antibodies Anti cd3 Cd25 Clone Cy5 5 Fitc Frozen Hnk 1 Human

Corresponding Organization : Icahn School of Medicine at Mount Sinai

Other organizations : Nantes Université, Inserm

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Variable analysis

independent variables

Anti-human antibodies: FITC-anti-CD127, PE-anti-CD3, PerCP-Cy5.5-anti-CD57, PE-Cy7-anti-CD4, APC-anti-CD25, APC-Cy7-anti-PD-1, BV510-anti-CD8

dependent variables

Outcomes measured from flow cytometry analysis

control variables

Freshly frozen PBMC from CTOT-09 study
Flow cytometry data acquisition (> 1 × 10^6 events) on a 3-laser FACSLyric flow cytometer
Data analysis using FlowJo software

controls

No positive or negative controls were explicitly mentioned in the provided information.

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