Flow cytometry was used to determine myeloid, macrophage, and microglia populations in single-cell preparations of the meninges, brainstem, hypothalamus, and thalamus in complete media: Iscove Modified Dulbecco Media (Life Technologies/Invitrogen, Carlsbad, CA), 10% Fetal Calf Serum, penicillin/streptomycin on ice. Samples were stained with an Ab mix (MACS buffer plus 2% inactivated rat serum), 2% anti-FcγII/III (clone 2.4G2), anti-CD11b (clone M1/70; BD HorizonTM), anti-CD45 (clone 30-F11; BD Pharmingen), anti-IL-13 (clone eBio13A; eBioscience), and IFN-γ (clone MG1.2; BD Pharmingen) for 45 min on ice, and then fixed in 2% paraformaldehyde before being permeabilized (saponin containing permeabilization buffer) for 1 h, at 4 °C. Samples were read using a BD FACS Fortessa machine (BD Biosciences, San Diego, CA), and data analysed by FlowJo (Tree Star, Ashland, OR) to be graphed with GraphPad Prism software.
Anti il 13 clone ebio13a
Manufactured by Thermo Fisher Scientific
Anti-IL-13 (clone eBio13A) is a monoclonal antibody that binds to the cytokine interleukin-13 (IL-13). It can be used for the detection and quantification of IL-13 in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Fortessa facs machine, by BD (1 mentions)
Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (1 mentions)
Anti cd45 clone 30 f11, by BD (1 mentions)
Brefeldin a, by Merck Group (1 mentions)
Anti il 4 clone 11b11, by Thermo Fisher Scientific (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
Facsaria flow cytometer, by BD (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Cytofix cytoperm solution, by BD (1 mentions)
Anti il17 clone ebio17b7, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti il 13 clone ebio13a
1
Immunophenotyping of Neuroimmune Populations
2
Multiparametric Flow Cytometry of Lymphocytes
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Single-cell suspensions were prepared from inguinal lymph node extracts. Surface antigen staining was performed using the following antibodies (eBioscience): anti-CD45 (clone 30-F11), anti-CD4 (clone RM4-5), anti-CD8 (clone 53–6.7), and anti-TCRγ (clone H57-597). For intracellular cytokine staining, isolated cells were activated in complete RPMI containing 0.5 µg ml−1 phorbol 12 myristate 13-acetate (PMA), 0.5 µg ml-1 ionomycin and 10 µg ml−1 brefeldin A (all from Sigma-Aldrich) for 4 h. After surface staining, cells were fixed and permeabilized using Cytofix/Cytoperm solution (BD). Cells were washed in perm/wash buffer and incubated for 30 min at 4°C with different antibody combinations specific for murine cytokines (all from eBioscience) diluted in perm/wash buffer: anti-Il4 (clone 11B11), anti–IL-13 (clone eBio13A), anti-Il17 (clone eBio17B7), and TNF (clone MP6-XT22). Immunostained cells were analyzed by flow cytometry using a FACS Aria flow cytometer (BD).
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