MBTP 24: Immunophenotyping of innate lymphoid cell 2 (ILC2) subsets from mouse mesentery and small intestine using flow cytometry

Miltenyi Biotec-tested panel 24 (MBTP 24)

This application protocol describes the flow cytometric analysis of ILC2 subsets after dissociation of mesentery and small intestine from healthy C57BL/6 mice. Mouse mesentery and small intestine are easily dissociated using the gentleMACS™ Dissociators to quickly obtain viable single cell suspensions ready for downstream applications, such as flow cytometric analysis.

Protocol

Gating strategy showing the analysis of ILC2 subsets from mouse mesentery and small intestine. Mesentery and small intestine cells were stained with the previously described panel to analyze ILC2 subsets. Samples were gated on lymphocytes using FSC-A/SSC-A gating (A). Samples were gated on single cells using FSC-W/FSC-H and SSC-W/SSC-H gating within the lymphocyte gate (data not shown). Live lymphocytes were defined as propidium iodide-negative and CD45.2+ cells (B). Within lineage-negative cells, ILC2s were identified as CD90.2+ Sca-1+ KLRG1+ cells (C, D). Alternative REAfinity™ Antibodies with similar performance results are available. Please refer to table 2 for more information.

Materials

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