Anti cd45ro pacific blue

Manufactured by BioLegend

Anti-CD45RO Pacific Blue is a laboratory reagent used for the identification and enumeration of CD45RO-expressing cells in flow cytometry applications. It binds specifically to the CD45RO surface antigen, which is expressed on memory T cells.

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2 protocols using anti cd45ro pacific blue

Multiparametric Flow Cytometry Panel

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The following antibodies were used for flow cytometric stainings anti-TACI PE (clone 1A1), anti-CD19 APC-Cy7, anti-CD27 PerCP-Cy5.5 or APC, anti-CD10 PE-Cy7, anti-IgM FITC, anti-CD21 APC, anti-CD69 PE-Cy7, anti-CD86 PE, anti-CD4 APC-Cy7, anti-CD25 PECy7, anti-CD127 PerCP-Cy5.5, anti-CD45RO Pacific Blue, anti-CXCR5 PerCP-Cy5.5, anti-PD-1 PE-Cy7, anti-CD25 PE, anti-CD25 PE-Cy7 (all from BioLegend, San Diego, Calif), anti-CD3 eFluor 605NC, anti-CD21 BD Horizon V450 (Becton Dickinson) and goat polyclonal anti-TACI biotin (R&D Systems). Intracellular staining with anti-Foxp3 Alexa Fluor 488 and anti-BCL6 PE (eBioscience, San Diego, Calif) was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience), in accordance with the manufacturer's instructions.

Romberg N., Virdee M., Chamberlain N., Oe T., Schickel J.N., Perkins T., Cantaert T., Rachid R., Rosengren S., Palazzo R., Geha R., Cunningham-Rundles C, & Meffre E. (2015). TNFRSF13B hemizygosity reveals TACI haploinsufficiency at later stages of B-cell development. The Journal of allergy and clinical immunology, 136(5), 1315-1325.

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Intracellular Cytokine Staining of PBMC

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Isolated 10⁶ PBMC were stimulated with phorbol myristate acetate (PMA) (25 ng/ml) and ionomycin (1 μg/ml) for 6 h at 37°C in RPMI‐1640 medium containing 10% FCS and 10 μg/ml brefeldin A. Immediately after stimulation, the cells were stained with anti‐CD3‐APC/Cy7 (BioLegend; clone HIT3a), anti‐CD4‐PerCP (BioLegend; clone RPA‐T4), anti‐CD8‐PE/Cy7 (eBioscience; clone RPA‐T8) and anti‐CD45RO‐Pacific Blue (BioLegend; clone UCHL1). Subsequently, cells were fixed and permeabilized using Cytofix/Cytoperm (BD), washed and stained for intracellular IL‐2 in Perm buffer with anti‐IL‐2‐APC [eBioscience; clone MQ1–17H12, in parallel with interferon (IFN)‐γ and tumour necrosis factor (TNF)‐α], assessed cytometrically with a fluorescence activated cell sorter (FACS)Canto II cytometer and analysed using FlowJo software (TreeStar, Inc.).

Costa N., Marques O., Godinho S.I., Carvalho C., Leal B., Figueiredo A.M., Vasconcelos C., Marinho A., Moraes‐Fontes M.F., Gomes da Costa A., Ponte C., Campanilho‐Marques R., Cóias T., Martins A.R., Viana J.F., Lima M., Martins B, & Fesel C. (2017). Two separate effects contribute to regulatory T cell defect in systemic lupus erythematosus patients and their unaffected relatives. Clinical and Experimental Immunology, 189(3), 318-330.

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