BM cells collected from 6- to 8-week-old C57BL/6 mice tibia and femur were flushed with PBS. Red blood cell (RBC) lysis buffer (BD Biosciences; CA, USA) was used to lyse red blood cells. Cells (2 × 106) were cultured at 37°C and 5% CO2 for 7 days in 6 cm dishes containing 3 ml Roswell Park Memorial Institute (RPMI)-1640 with 10% FBS, 1% streptomycin and penicillin, 1% MEM nonessential amino acids (NEAA) solution, 1% sodium pyruvate (Gibco, NY, USA), 2 μl 2-mercaptoethanol (Sigma-Aldrich, St. Louis, USA), 50 ng/ml GM-CSF (PeproTech), and 40 ng/ml interleukin (IL)-6 (PeproTech). The cells were harvested for testing or sorting by FACS Aria III (BD Biosciences, CA, USA) using CD11b-fluorescein isothiocyanate (FITC) (eBioscience, CA, USA) and Gr-1-PerCP Cy5.5 (eBioscience). Flow cytometry verified that all isolated MDSCs yielded >90% pure population. The purified MDSCs were then adoptively transferred to mice via the tail vein just before UUO. For cell tracing, some UUO mice were injected with MDSCs and stained with CFSE (Invitrogen).
Cd11b fluorescein isothiocyanate fitc
Manufactured by Thermo Fisher Scientific
Sourced in United States
CD11b-fluorescein isothiocyanate (FITC) is a fluorescently labeled antibody that binds to the CD11b antigen, which is expressed on the surface of various immune cells, including monocytes, macrophages, and neutrophils. The FITC fluorescent dye is attached to the antibody, allowing for the detection and analysis of CD11b-positive cells using flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
Rbc lysis buffer, by BD (1 mentions)
Interleukin il 6, by Thermo Fisher Scientific (1 mentions)
Gr 1 percp cy5, by Thermo Fisher Scientific (1 mentions)
Facsaria 3, by BD (1 mentions)
Mem non essential amino acid, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
Gm csf, by Thermo Fisher Scientific (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Facsaria 2, by BD (1 mentions)
Facscelesta, by BD (1 mentions)
F4 80 pe, by Thermo Fisher Scientific (1 mentions)
2 protocols using cd11b fluorescein isothiocyanate fitc
1
Murine MDSC Isolation and Adoptive Transfer
2
Phenotypic Characterization of Immune Cells
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Phenotype analysis of the macrophages and lymphocytes was executed employing a BD FACS AriaII flow cytometer (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), as previously detailed [21 (link)]. In brief, the cells were labeled with CD11b-fluorescein isothiocyanate (FITC) (11-0112-82; eBioscience, San Diego, CA, USA), CD206-allophycocyanin (17-2061-82; eBioscience), IL-17-phycoerythrin (PE) (12-7177-81; eBioscience), and F4/80-PE (12-4801-82; eBioscience), according to the manufacturer's recommended protocol. For the assessment of M2 macrophages prevalence, the cells were subjected to incubation with PE-conjugated F4/80 antibodies or allophycocyanin-conjugated CD206. For the analysis of Th17 cell prevalence, the cells were incubated with FITC-conjugated CD4 antibodies or PE-conjugated IL-17 antibodies. Flow cytometry was performed on BD FACSCelesta (BD Biosciences, Franklin Lakes, NJ, USA), and data were analyzed by FlowJo software (FlowJo v10).
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