Cd45ra viogreen

Manufactured by Miltenyi Biotec

CD45RA-VioGreen is a fluorochrome-conjugated antibody that binds to the CD45RA antigen, which is expressed on a subset of T cells. It is a lab equipment product intended for use in flow cytometry applications.

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2 protocols using cd45ra viogreen

Immunophenotyping of Expanded UCB Cells

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The antigenic phenotype of UCB cells was assessed before and during ex vivo expansion. Approximately 1 × 10⁵ cells per test were incubated with the required antibody panel as per manufacturer’s instructions for 30 min at room temperature. All antibodies were preconjugated and monoclonal. Progenitor panel: CD34-PerCP-Vio700, CD38-FITC, CD45RA-VioGreen, CD90-APC-Vio770, CD133-APC, CD135-PE-Vio770 (Miltenyi Biotech) and CD33-PE (BD Biosciences). Mature stage myeloid panel: CD13-APC, CD14-PE, CD15-VioGreen, CD34-PerCP-Vio700, CD33-APC-Vio770, CD38-FITC, CD123-VioBlue (Miltenyi Biotech). Analysis was conducted with CITRUS (cluster identification, characterisation and regression) software [31 (link)]. The software uses a regularized supervised learning algorithm to determine the populations and features of the samples being analysed in a correlative manner. The results of a CITRUS run are clusters (populations) that differentiate the observed endpoint of the samples, and the features (relative population abundance or median expression of a functional marker) of the clusters that are responsible. All CITRUS analyses were conducted using a Significance Analysis of Microarrays (SAM) association model and a minimum cluster size of 4%, at least 5000 events from each sample were included for the analysis.

Moore R.L., Worrallo M.J., Mitchell P.D., Harriman J., Glen K.E, & Thomas R.J. (2017). Immobilisation of Delta-like 1 ligand for the scalable and controlled manufacture of hematopoietic progenitor cells in a stirred bioreactor. BMC Biotechnology, 17, 65.

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Phenotyping of T and NK cell subsets

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T cells and NK cells were obtained as described above. Cells were incubated with fluorochrome-labeled antibodies at 4°C. Specific antibodies for the T cell subsets include CD3 V450, CCR7 FITC, CD45RO APC and CD45RA Viogreen (Miltenyi Biotech, Auburn, CA). Specific antibodies for the NK cell subsets include, CD56 FITC, CD11b V450, CD3 Viogreen, and CD27 APC (Miltenyi Biotech, Auburn, CA). Flow cytometry was performed as previously described (19 (link)).

del Campo S.E., Levine K.M., Mundy-Bosse B.L., Grignol V.P., Fairchild E.T., Campbell A.R., Trikha P., Mace T.A., Paul B.K., Jaime-Ramirez A.C., Markowitz J., Kondadasula S.V., Guenterberg K.D., McClory S., Karpa V.I., Pan X., Olencki T.E., Monk J.P., Mortazavi A., Tridandapani S., Lesinski G.B., Byrd J.C., Caligiuri M.A., Shah M.H, & Carson WE I.I.I. (2015). The Raf Kinase Inhibitor Sorafenib Inhibits JAK-STAT Signal Transduction in Human Immune Cells. Journal of immunology (Baltimore, Md. : 1950), 195(5), 1995-2005.

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