At the time of euthanasia, one inch pieces of ileum and colon were collected, cleaned of any luminal contents and ~50 mg tissue piece was enzymatically digested to prepare single cell suspension as described earlier (Bie et al. 2011 ). Isolated cells were re‐suspended in FACS buffer containing Fc block and incubated with fluorescently labeled antibodies for 20 min at 4°C. After washing, 50,000 FITC‐labeled counting beads were added to each sample and analyzed by flow cytometry (BD Biosciences, Canto II) with stopping gate set to count 10,000 beads. The data were analyzed using FlowJO (Tree Star Inc.) software. The following antibodies were used: Antimouse CD45‐PE (leukocytes), antimouse CD11b‐PerCP‐Cy5.5 (macrophages) antimouse Ly6C‐APC and the respective isotype controls (all antibodies were obtained from eBiosciences).
Antimouse cd45 pe
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Antimouse CD45-PE is a fluorescently labeled antibody that binds to the CD45 antigen expressed on the surface of mouse leukocytes. It is designed for use in flow cytometry applications to identify and quantify different subsets of mouse immune cells.
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Lab products found in correlation
Anti mouse ly6c apc, by Thermo Fisher Scientific (1 mentions)
Anti mouse cd11b percp cy5, by Thermo Fisher Scientific (1 mentions)
Canto 2, by BD (1 mentions)
Ab32529, by Abcam (1 mentions)
Ab87540, by Abcam (1 mentions)
Ab91109, by Abcam (1 mentions)
Anti mouse cd62l, by BioLegend (1 mentions)
Anti mouse cd44 pe, by Thermo Fisher Scientific (1 mentions)
Apc anti mouse cd8a, by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
2 protocols using antimouse cd45 pe
1
Flow Cytometric Analysis of Intestinal Immune Cells
2
CD8+ Tem Cell Phenotyping by Flow Cytometry
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We used flow cytometry to isolate and count CD8+ Tem cells. Cells were labeled with the following fluorescently conjugated monoclonal antibodies: anti-mouse CD45-PE (12045181, eBioscience, San Diego, CA, USA), anti-mouse CD8a-APC (17008181, eBioscience), anti-mouse CD44-PE (25044181, eBioscience), and anti-mouse CD62L (104432, BioLegend, San Diego, CA, USA). After the CD8+ Tem cells were sorted by flow cytometry, they were stained with antibodies against mTOR (ab87540, Abcam, Cambridge, MA, USA), interferon γ (IFN-γ) (11731181, eBioscience), T-bet (ab91109, Abcam), S6K (ab32529, Abcam) and Eomes (53-4875-82, eBioscience).
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