Lymph nodes (axillary or popliteal) were collected in FACS buffer (PBS1X, 10%FCS, 2mM EDTA) on ice. Single-cell suspensions were obtained by mechanical disruption through a 70-mm cell strainer (BD Biosciences). Erythrocytes were lysed with 1 ml of ACK lysing buffer (Gibco). After incubation with 5 μg/ml of anti-CD16/32 (rat mAb 2.4G2, mouse Fc block BD) for 15 min at 4°C, cell surface antigens were stained for 30 min at 4°C. When a biotin antibody was used, the cells were then incubated with a streptavidin-fluorophore conjugate for 20 min at 4°C. Flow cytometric analysis was performed on a BD LSRFortessa. Antibodies used: from BD; Anti-IgM-e710 (R6-60.2, 550881), Anti-IgD-BV786 (11-26c.2a, 563618), Anti- T-and B-cell activation antigen-FTIC (GL7, 553666), Anti-Igλ1,2,3-BV650 (R26-46, 744526), Anti-CD95-PE-Cy7 (Jo2, 557653) streptavidin-BV711 (563262), streptavidin-AF647, from Biolegend; Anti-CD38-PB (90, 102719), Anti-CD138-BV421 (281-2, 142508), Anti-B220-BV605 (RA3-6B2, 103244) live/dead marker Zombie NIR (Biolegend, 423106) and from eBiosciences; Anti-T-and B-cell activation antigen-e660 (GL7, 50-5902-82), Anti-CD4-eF780 (RM4-5, 47-0042-82), Anti-CD8-eF780 (53-6.7, 47-0081-82), Anti-NK1.1-eF780 (PK136, 47-5941-82), Anti-F4/80-eF780 (BM8, 47-4801-82), Anti-TM4-Core-biot (5μg/mL) provided by A.T.M and L.S (Fred Hutchinson Cancer Research Center, Seattle).
Anti f4 80 ef780
Manufactured by Thermo Fisher Scientific
The Anti-F4/80-eF780 is a fluorescent-conjugated antibody that binds to the mouse F4/80 antigen, a glycoprotein expressed on the surface of macrophages and microglia. This product is designed for use in flow cytometry and other immunological applications to identify and study macrophage populations.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd95 pecy7, by BioLegend (2 mentions)
70 mm cell strainer, by BD (2 mentions)
Ack lysing buffer, by Thermo Fisher Scientific (2 mentions)
Lsrfortessa, by BD (2 mentions)
Anti cd4 ef780, by Thermo Fisher Scientific (2 mentions)
Anti cd8 ef780, by Thermo Fisher Scientific (2 mentions)
Anti nk1.1 ef780, by Thermo Fisher Scientific (2 mentions)
Streptavidin af647, by BioLegend (1 mentions)
Anti b220 bv605, by BioLegend (1 mentions)
Anti igd bv786, by BioLegend (1 mentions)
Anti cd38 pb, by BioLegend (1 mentions)
Anti cd138 bv421, by BioLegend (1 mentions)
Anti cd16 32, by BD (1 mentions)
Streptavidin bv711, by BioLegend (1 mentions)
2 protocols using anti f4 80 ef780
1
Phenotyping Lymphocyte Populations by Flow Cytometry
2
Multiparameter Flow Cytometry of Lymph Nodes
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Lymph nodes (axillary or popliteal) were collected in FACS buffer (1× PBS, 10% FCS, 2 mM EDTA) on ice. Single-cell suspensions were obtained by mechanical disruption through a 70-mm cell strainer (BD Biosciences). Erythrocytes were lysed with 1 ml ACK lysing buffer (Gibco). After incubation with 5 µg/ml anti-CD16/32 (rat mAb 2.4G2, mouse Fc block; BD Biosciences) for 15 min at 4°C, cells were stained for 30 min at 4°C. When a biotinylated antibody was used, the cells were then incubated with a streptavidin–fluorophore conjugate for 20 min at 4°C. Flow cytometric analysis was performed on a BD LSRFortessa and Symphony. Antibodies used: from BD Biosciences, anti-IgM-e710 (R6-60.2, 550881), anti-IgD-BV786 (11-26c.2a, 563618), anti-CD95-PE-Cy7 (Jo2, 557653), and streptavidin-BV711 (563262); from BioLegend, anti-CD38-PB (90, 102719), anti-B220-BV605 (RA3-6B2, 103244), and live/dead marker Zombie NIR (423106); and from eBiosciences, anti-T and -B cell activation antigen-e660 (GL7, 50-5902-82), anti-CD4-eF780 (RM4-5, 47-0042-82), anti-CD8-eF780 (53-6.7, 47-0081-82), anti-NK1.1-eF780 (PK136, 47-5941-82), anti-F4/80-eF780 (BM8, 47-4801-82), and anti-TM4-Core-biot (5 µg/ml) provided by Andrew T. McGuire and L. Stamatatos.
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