Alexafluor488 ly6c

Manufactured by BioLegend

Sourced in United States

AlexaFluor488-Ly6C is a fluorescently-labeled antibody reagent produced by BioLegend. It recognizes the Ly6C antigen, which is expressed on various cell types. The AlexaFluor488 fluorophore is conjugated to the antibody, allowing for detection and analysis of Ly6C-positive cells using flow cytometry or other fluorescence-based techniques.

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2 protocols using alexafluor488 ly6c

Quantification of GM-CSF Receptor Expression in Mouse Immune Cells

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GM‐CSFRα and β expression was quantified in mouse BALF, blood and bone marrow neutrophils by flow cytometry and expressed as the MFI geometric mean. Cells were stained for GM‐CSFRα with APC‐CAM‐3003 (labelled with Lightning‐Link APC as per the manufacturer's instructions, Innova Biosciences, Cambridge, UK), GM‐CSFRβ with PE‐CD131 (JORO50, BD Pharmingen), and BUV395‐CD45 (4 μg/ml, clone 30‐F11, BD Horizon), BV421‐Ly6G (12 μg/ml, clone 1A8, BD Horizon), AlexaFluor488‐Ly6C (12 μg/ml, clone HK1.4, Biolegend, San Diego, California, USA), AlexaFluor488‐CD11b (12 μg/ml, clone M1/70, BD Pharmingen) and eF780 viability dye (1:1000 dilution, eBioscience). All flow cytometry was performed on one day on a single instrument for each study.

De Alessandris S., Ferguson G.J., Dodd A.J., Juss J.K., Devaprasad A., Piper S., Wyatt O., Killick H., Corkill D.J., Cohen E.S., Pandit A., Radstake T.R., Simmonds R., Condliffe A.M., Sleeman M.A., Cowburn A.S., Finch D.K, & Chilvers E.R. (2019). Neutrophil GM‐CSF receptor dynamics in acute lung injury. Journal of Leukocyte Biology, 105(6), 1183-1194.

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Multiparametric Flow Cytometry Analysis

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Surface and intracellular staining were performed as described previously using the indicated antibodies [33 (link)]. From BioLegend: Alexa fluor 488-Ly6C, PE-CD3, PE-CD86, APC-CD80, APC-I-A/I-F, Biotin-ICAM I, Biotin-H2kb, Alexa fluor 488-IFN-γ, Biotin-PD-L1, PE-Cy7-Ly6G, PE-CD34, PE-CD14, PE-F4/80, PE-Cy7-streptavidin, APC-streptatividin; From eBioscience: PE-CD4, FITC-IA/IE; From Raybiotech: FITC-class I H-2Kb; From BD bioscience pharmigen: v500-CD4, PE-CD8alpha, APC-CD11b, AF647-Ki67; from Southernbiotech: from Invitrogen: APC-CD11c; PE-streptavidin, FITC-streptavidin; from AbD Serotec: PE-CD62L; from R&D Systems: anti-human/mouse PE-Arginase 1. Viability was established by flow cytometry using the Fixable viability dye-eF780 from eBioscience. From Santa Cruz Biotechnology, NOS2-PE. When indicated, cells samples were treated overnight with 100 ng/ml lipololysaccharide (LPS) from Salmonella enterica serotype abortusequi (SIGMA).

Liechtenstein T., Perez-Janices N., Gato M., Caliendo F., Kochan G., Blanco-Luquin I., Van der Jeught K., Arce F., Guerrero-Setas D., Fernandez-Irigoyen J., Santamaria E., Breckpot K, & Escors D. (2014). A highly efficient tumor-infiltrating MDSC differentiation system for discovery of anti-neoplastic targets, which circumvents the need for tumor establishment in mice. Oncotarget, 5(17), 7843-7857.

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