For study of the myeloid population, mice were injected subcutaneously with 0.5 × 106 CT26 colon carcinoma cells per mouse. Mice were sacrificed at tumor size of 1 cm. Tumors were disaggregated using collagenase and red blood cells were lysed using RBC lysis buffer. Cells were Fc blocked and stained with fluorochrome-conjugated antibodies for 30 min. The live/dead stain used was Zombie/NIR (Biolegend). For intracellular staining cells were first permeabilised using Foxp3 Transcription Factor Fixation/Permeabilization Concentrate and Diluent solution (eBioscience). The flourochrome-conjugated antibodies used were, anti-NOS2 APC (clone CXNFT) from eBioscience, anti CD45 BV711 (clone 30-F11), anti CD11c (clone N418), anti CD11b (clone M1/70), anti CD274 BV421 (clone 10F.9G2), anti CD80 BV650 (clone 16–10A1) from Biolegend, anti Arg1 Fluorescein from R&D systems. Acquisition was performed on an LSR Fortessa SORP HTS flow cytometer. Data analysis was performed using FlowJo 10.
Foxp3 transcription factor fixation permeabilization concentrate and diluent solution
Manufactured by Thermo Fisher Scientific
The Foxp3 Transcription Factor Fixation/Permeabilization Concentrate and Diluent solution is a kit designed for the intracellular staining and detection of the Foxp3 transcription factor. The concentrate solution is used for the fixation and permeabilization of cells, while the diluent solution is used for the dilution of the concentrate. This kit provides the necessary reagents for the preparation of samples for flow cytometric analysis of Foxp3-expressing cells.
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Lab products found in correlation
2 protocols using foxp3 transcription factor fixation permeabilization concentrate and diluent solution
1
Immunophenotyping Myeloid Cells in Tumor
2
Intracellular Cytokine Profiling by Flow
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For the analysis of intracellular IFNγ, IL-10, IL-17A, and IL-17F, cells were stimulated for 4 h in IMDM containing 50 ng/ml PMA, 500 ng/ml ionomycin, and 10 µg/ml brefeldin A. After surface staining, the cells were fixed, permeabilized, and intracellularly stained with the FOXP3/Transcription Factor Fixation/Permeabilization Concentrate and Diluent solution (e-bioscience) and antibody, according to the manufacturer’s protocol. Data were collected by flow cytometry.
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