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Fitc conjugated f4 80 antibody sc 71085

Manufactured by Santa Cruz Biotechnology
Sourced in United States

The FITC-conjugated F4/80 antibody (SC-71085) from Santa Cruz Biotechnology is a research-use antibody conjugated with the fluorescent dye FITC (Fluorescein isothiocyanate). The F4/80 antigen is a member of the EGF-TM7 family of proteins and is expressed on the surface of mature mouse macrophages. This antibody can be used to detect and analyze macrophages in various experimental applications.

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2 protocols using fitc conjugated f4 80 antibody sc 71085

1

Quantifying M2 Macrophage Phenotypes

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The phenotype of macrophages was identified by analyzing the surface markers of macrophages using flow cytometer. Briefly, macrophages were harvested, washed, and resuspended in 100 µl PBS solution at a density of 5 × 106 cells/ml. For Thp1 macrophages, cells were incubated with FITC-conjugated F4/80 antibody (SC-71085, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and PE-conjugated CD163 antibody (12-1639-42, eBioscience, San Diego, CA, USA) for 30 mins at 37°C. For Raw264.7 macrophages, cells were incubated with FITC-conjugated F4/80 antibody (SC-71085, Santa Cruz) and PE-conjugated CD206 antibody (141705, Biolegend, San Diego, CA, USA) for 30 mins at 37°C. For phenotype analysis of primary macrophages extracted from mouse 4T1-Luc xenografts, cells were incubated with CD45-PE-Cy7 (25-0451-82, eBioscience), FITC-conjugated F4/80 antibody (SC-71085, Santa Cruz) and PE-conjugated CD206 antibody (141705, Biolegend) for 30 mins at 37°C. After incubation, cells were washed once with PBS and subjected to analysis using a FC500 flow cytometry (Beckman Coulter, Fullerton, CA, USA) or a FACSAria III flow cytometer (BD Biosciences, San Diego, CA, USA). The F4/80+/CD163+ subpopulation, the F4/80+/CD206+ subpopulation or the CD45+/F4/80+/CD206+ subpopulation were quantified and defined as M2 phenotype macrophages.
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2

Macrophage Phenotyping by Flow Cytometry

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For the phenotype analysis of Raw264.7 macrophages, cells were first treated as indicated. Then, Raw264.7 cells were harvested and incubated with FITC-conjugated F4/80 antibody (SC-71085, Santa Cruz, CA, USA), PE-conjugated CD206 antibody (141,705, Biolegend, CA, USA), or PE-Cy7-conjugated CD206 antibody (E-AB-F1135H, Elabscience, Houston, TX, USA) for 30 min at 37 °C. For the phenotype analysis of THP1 macrophages, cells were treated as indicated, harvested and incubated with APC-conjugated F4/80 antibody (17-4801-82, Invitrogen) and PE-conjugated CD163 antibody (12-1639-42, eBioscience) for 30 min at 37 °C. For the phenotypic analyses of primary macrophages isolated from mouse 4 T1-Luc xenografts, cells were incubated with CD45-PE-Cy7 (25-0451-82, eBioscience), F4/80-APC antibody (17-4801-82, Invitrogen), and CD206-PE antibody (141,705, Biolegend) for 30 min at 37 °C. For PD-L1 expression analyses of primary macrophages, cells were incubated with CD45-PE-Cy7 (25-0451-82, eBioscience), F4/80-FITC antibody (SC-71085, Santa Cruz), and PD-L1-APC antibody (124,312, Biolegend) for 30 min at 37 °C. After incubation, the cells were washed once with PBS and subjected to flow cytometry analysis.
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