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Cd45ro pe cy7

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The CD45RO-PE-Cy7 is a fluorescently labeled antibody that binds to the CD45RO antigen expressed on the surface of certain human immune cells. It is used as a tool in flow cytometry applications to identify and analyze specific cell populations within a sample.

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Lab products found in correlation

Ultracomp ebeads, by Thermo Fisher Scientific (1 mentions) Cd3 apc, by Thermo Fisher Scientific (1 mentions) Cd4 pe, by Thermo Fisher Scientific (1 mentions) Ccr7 bv421, by BD (1 mentions) Cd8a apc efluor 780, by Thermo Fisher Scientific (1 mentions) Cd56 pe, by Thermo Fisher Scientific (1 mentions) 35 m strainer, by BD (1 mentions) Cd19 pe, by BD (1 mentions) Cd27 apc efluor780, by Thermo Fisher Scientific (1 mentions) Cxcr5 apc, by BD (1 mentions) Cd3 percp cy5, by BD (1 mentions) Cd8 bv650, by BioLegend (1 mentions) Cd4 bv605, by BioLegend (1 mentions) Fixable viability dye efluor 506, by Thermo Fisher Scientific (1 mentions) Lsr 2, by BD (1 mentions) Cd8 apc efluor 780, by Thermo Fisher Scientific (1 mentions) Cd4 apc efluor 780, by Thermo Fisher Scientific (1 mentions) Cd45ra percp cy5, by Thermo Fisher Scientific (1 mentions) Cd3 pe, by Thermo Fisher Scientific (1 mentions) Easysep, by STEMCELL (1 mentions)

3 protocols using cd45ro pe cy7

1

Isolation and Characterization of CD20+ and CD20- Cells

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Buffy coat samples were washed with PBS 2% FCS and stained for 60 min at 4 °C with Rituximab CD20-FITC, CD3-APC (eBioscience), CD8a -APC-eFluor780 (eBioscience), CD45RO- PE-Cy7 (eBioscience), CCR7-BV421 (BD), and markers CD4-PE (eBioscience), CD56-PE (eBioscience) and CD19-PE (BD). Cells were washed and filtered using a 35 µm strainer (Falcon). Propidium Iodide (1 µg/ml) was used for exclusion of dead cells. From the CD3 + CD8 + CD45RO + CCR7- population, 1000 live CD20 + and 1000 live CD20-cells were sorted using a Beckman Coulter MoFlo Astrios cytometer. UltraComp eBeads (Thermo Fisher Scientific) were used as compensation controls.
Vlaming M., Bilemjian V., Freile J.Á., Lourens H.J., van Rooij N., Huls G., van Meerten T., de Bruyn M, & Bremer E. (2021). CD20 positive CD8 T cells are a unique and transcriptionally-distinct subset of T cells with distinct transmigration properties. Scientific Reports, 11, 20499.
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2

TFH Cell Identification by Flow Cytometry

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The following antibodies were used: CD8 BV650, CD4 BV605, and CD57 PacBlue all obtained from Biolegend. CXCR5-APC and CD3-PercpCy5.5 were obtained from Becton Dickinson. CD27 APC-eFLuor 780, CD45RO PeCy7, and Fixable Viability Dye eFluor 506 were all obtained from eBiosciences. Flow cytometry was done on a BD LSRII and data analyzed using FlowJo (Treestar). To identify TFH cells, samples were gated on live cells within the lymphocyte gate after doublet discrimination. TFH cells were then defined by gating on CD3+CD8−CD4+CXCR5+ cells.
Knorr D.A., Wang H., Aurora M., MacMillan M.L., Holton S.G., Bergerson R., Cao Q., Weisdorf D.J., Cooley S., Brunstein C., Miller J.S., Wagner J.E., Blazar B.R, & Verneris M.R. (2016). Loss of T Follicular Helper Cells In the Peripheral Blood Of Patients With Chronic Graft vs. Host Disease. Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation, 22(5), 825-833.
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3

Isolation of Naive T-Cell Subpopulations

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Human peripheral blood mononuclear cells were isolated from fresh whole blood by density gradient centrifugation using Histopaque (ρ = 1.077 g/cm 3 , Sigma, USA). The subpopulations of nTh (CD4 + CD45RO -) and nCTL (CD8 + CD45RO -) were isolated by negative magnetic immunoseparation using commercial kits series EasySep (Stemcell Technologies, UK). The purity of isolated cells was monitored by flow cytometry using a panel of fluorescent-labeled mAb: CD3-PE, CD45RO-PE-Cy7, CD45RA-PerCP-Cy5.5, and CD8-APC-eFluor780 (or CD4-APC-eFluor780) (eBioscience, USA). We verified that the subpopulations of nTh and nCTL T-cells were purified to more than 98% (Figure 1).
Filatova E.N., Anisenkova E.V., Presnyakova N.B, & Utkin O.V. (2016). DR3 regulation of apoptosis of naive T-lymphocytes in children with acute infectious mononucleosis. Acta microbiologica et immunologica Hungarica, 63(3).
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