Monocytes were isolated from normal peripheral blood mononuclear cells (PBMC) and differentiated for 7 days. Briefly, CD14+ monocytes were purified from PBMC by positive selection using anti-CD14 monoclonal antibody-coated magnetic microbeads (Miltenyi Biotech) and differentiated in the presence of M-CSF and GM-CSF as described previously [23 (link)]. MDMs were infected with a multiplicity of infection (MOI) of 0.5. Mock infection was performed using equal amount of HEK293T supernatant. MDM supernatants were collected 10 to 12 days post-infection.
Anti cd14 monoclonal antibody coated magnetic microbeads
Manufactured by Miltenyi Biotec
Anti-CD14 monoclonal antibody-coated magnetic microbeads are designed for the isolation and enrichment of CD14-positive cells from complex biological samples. The microbeads are superparamagnetic and coated with monoclonal antibodies specific for the CD14 cell surface marker.
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Lab products found in correlation
3 protocols using anti cd14 monoclonal antibody coated magnetic microbeads
1
Differentiation and Infection of Monocyte-Derived Macrophages
2
Monocyte-derived Macrophage Generation
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Monocytes-derived macrophages (MDMs) were generated from normal peripheral blood mononuclear cells (PBMC). PBMCs from healthy donor were isolated by Ficoll-Hypaque gradient centrifugation. CD14 + monocytes were purified by positive selection using anti-CD14 monoclonal antibody-coated magnetic microbeads (Miltenyi Biotech, Auburn, CA) and differentiated in presence of 1 pg/ml M-CSF and 1 × 106 IU/ml GM-CSF (R&D Systems) as described previously [25 (link)]. Half the volume of media was replaced every third day with fresh media containing GM-CSF and M-CSF for 7–8 days to differentiate them into MDMs.
3
Generation of Monocyte-Derived Macrophages
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+ Expand
Monocytes-derived macrophages (MDMs) were generated from normal peripheral blood mononuclear cells (PBMC). PBMCs from healthy donor were isolated by Ficoll-Hypaque gradient centrifugation. CD14 + monocytes were purified by positive selection using anti-CD14 monoclonal antibody-coated magnetic microbeads (Miltenyi Biotech, Auburn, CA) and differentiated in presence of 1 pg/ml M-CSF and 1 × 106 IU/ml GM-CSF (R&D Systems) as described previously [25 (link)]. Half the volume of media was replaced every third day with fresh media containing GM-CSF and M-CSF for 7–8 days to differentiate them into MDMs.
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