1 × 106 cell suspension from lung digests and BAL were incubated with antibody cocktail containing APC-anti-CD206 (BioLegend. San Diego, CA), PerC-Cy5-anti-F4/80, PE-Cy7-anti-Ly6G, PE-Cy3-anti-p-STAT6, APC-Cy7-anti-CD11b, BV421-anti-Siglec-F (BD Biosciences, Franklin Lakes, NJ and eBioscience, San Diego, CA). After incubation at dark for 40 min, the stained cells were washed for 2 times and analyzed on FACScan cytometer and Flow Jo software, version 8.8.4 (Becton, Dickinson and Company, Franklin Lakes, NJ).
Perc cy5 anti f4 80
Manufactured by BD
Sourced in United States
PerC-Cy5-anti-F4/80 is a fluorescently-labeled antibody that targets the F4/80 antigen found on mouse macrophages. It is used for the identification and analysis of macrophages in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti ly6g pe cy7, by BD (3 mentions)
Bv421 anti siglec f, by BD (3 mentions)
Anti cd11b apc cy7, by BD (3 mentions)
Facscan cytometer, by BD (2 mentions)
Pe cy3 anti p stat6, by BD (2 mentions)
Anti cd206 apc, by BioLegend (2 mentions)
Anti cd45 pe, by BD (1 mentions)
Facsaria 3 instrument, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Flow jo software version 8, by BD (1 mentions)
3 protocols using perc cy5 anti f4 80
1
Immunophenotyping of Lung Immune Cells
2
Immune Cell Profiling by Flow Cytometry
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The 0.3 × 106 cell suspensions of lung digests or BAL were incubated with an antibody cocktail containing PE-anti-CD45, PerC-Cy5-anti-F4/80, PE-Cy7-anti-Ly6G, APC-Cy7-anti-CD11b, and BV421-anti-Siglec-F (BD Biosciences, Franklin Lakes, NJ, USA; eBiosciences, San Diego, CA, USA). After incubation with the antibody cocktail for 40 min at room temperature, the cells were washed twice with PBS. The stained cells were analyzed by a BD FACSAria™ III instrument and BD FACSDiva™ software (BD Biosciences, San Jose, CA, USA). All data were analyzed using FlowJo software, version 8.8.4 (Tree Star Inc., Ashland, OR, USA).
3
Inflammatory Cytokine and Cell Profile
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The concentration of TNF-α, IL-6, IL-1β and Calreticulin in BAL or cell supernatants were measured by ELISA assay according to industrial instructions (R&D systems, Minneapolis, MN).
Flow Cytometry analysis 1 × 10 6 cell suspension from lung digests and BAL were incubated with antibody cocktail containing APCanti-CD206 (BioLegend. San Diego, CA), PerC-Cy5-anti-F4/80, PE-Cy7-anti-Ly6G, PE-Cy3-anti-p-STAT6, APC-Cy7-anti-CD11b, BV421-anti-Siglec F (BD Biosciences, Franklin Lakes, NJ and eBioscience, San Diego, CA). The cells stained with uorescence-minus-one (FMO) antibody cocktail were unstained controls. The cells were incubated with antibod cocktail in PBS supplied with 3% FBS for 40 min and washed with PBS for 2 times at room temperature. Analysis was performed on FACScan cytometer and all data were analyzed using FlowJo software, version 8.8.4 (Becton, Dickinson and Company, Franklin Lakes, NJ).
Flow Cytometry analysis 1 × 10 6 cell suspension from lung digests and BAL were incubated with antibody cocktail containing APCanti-CD206 (BioLegend. San Diego, CA), PerC-Cy5-anti-F4/80, PE-Cy7-anti-Ly6G, PE-Cy3-anti-p-STAT6, APC-Cy7-anti-CD11b, BV421-anti-Siglec F (BD Biosciences, Franklin Lakes, NJ and eBioscience, San Diego, CA). The cells stained with uorescence-minus-one (FMO) antibody cocktail were unstained controls. The cells were incubated with antibod cocktail in PBS supplied with 3% FBS for 40 min and washed with PBS for 2 times at room temperature. Analysis was performed on FACScan cytometer and all data were analyzed using FlowJo software, version 8.8.4 (Becton, Dickinson and Company, Franklin Lakes, NJ).
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