The peritoneal cells and cells from adipose tissues were collected as described previously. The cells were incubated with Fc Block CD16/32 antibodies (Thermo Fisher Scientific) followed by surface antibodies for 30 min on ice in the dark. RELMα, Ki67, NOS2, TNFα, and CD206 were stained using Foxp3 Transcription Factor Staining (Thermo Fisher Scientific) or BD Cytofix/Cytoperm™ Fixation/Permeabilization Kit (BD). The cells were stained with LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit (Thermo Fisher Scientific) to detect live cells. Flow Cytometry was performed on a BD LSRII, and results were analyzed by the FlowJo program. The following antibodies were used for flow cytometry analysis to detect each experiment. For peritoneal cells, PE-CD11b, eFluor450-F4/80, Alexa488-NOS2, PerCP-Cy5.5-Ki67, and Biotinylated RELMα with Alexa 647-Streptavidin were used. The following antibodies were used for flow cytometry analysis to detect Treg cells and macrophages: BV711-CD45, eFluor450-B220, FITC-CD25, PE-F4/80, eFluor450-F4/80, APC-eF780-CD11c, APC-Foxp3, PE-Cy7-CD206, PerCP-Cy5.5-CD11b, BV605-CD4, PE-NOS2, and Alexa488-TNFα.
Fc block cd16 32 antibodies
Manufactured by Thermo Fisher Scientific
Fc Block CD16/32 antibodies are a lab equipment product manufactured by Thermo Fisher Scientific. They are designed to block Fc receptors on cells, such as CD16 and CD32, to prevent non-specific binding of antibodies during immunoassays or cell-based experiments. This core function helps to minimize background noise and improve the specificity of experimental results.
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Lab products found in correlation
Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (2 mentions)
Cytofix cytoperm fixation permeabilization kit, by BD (1 mentions)
Lsr 2, by BD (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Dnase 1, by Roche (1 mentions)
Collagenase 1, by Worthington (1 mentions)
Ack lysing buffer, by Quality Biological (1 mentions)
2 protocols using fc block cd16 32 antibodies
1
Flow Cytometric Analysis of Peritoneal and Adipose Macrophages
2
Comprehensive Lung and Spleen Cell Isolation
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Lung was digested in RPMI (Thermo Fisher) with 0.5 mg/ml Collagenase I (Worthington) and 0.2 mg/ml DNase I (Roche) for 1 hr. Digested lung tissues were minced through 100 μm strainer. Spleen was directly minced through 100 μm strainer. Minced tissues were additionally filtered with 40 μm strainer after red blood cell lysis by ACK lysing buffer (Quality Biological). After incubation with Fc Block CD16/32 antibodies (Thermo Fisher Scientific), the cells from lung and spleen were further incubated with surface antibodies for 30 min on ice in the dark. Washed cells were stained with LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Thermo Fisher Scientific). BD LSRII was used for flow cytometry, and results were analyzed by FlowJo software. The following antibodies were used for flow cytometry analysis to detect CD4 T cell, CD8 T cell, γδ T cell, neutrophil, eosinophil, and macrophage: CD45-BV711, MerTK-FITC, CD64-BV605, F4/80-eFluor450, Ly6C-PerCP-Cy5.5, CD11c-APC, CD169-PE, CD86-PE-Cy7, CD3-BV605, CD4-PE-Cy7, CD8-eFluor450, TCR γ/δ-PE, Ly6G-APC, SiglecF-BV605, CD62L-PerCP-Cy5.5, CD44-APC-Cy7.
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