MBTP 13: Immunophenotyping of monocytes from mouse spleen using flow cytometry

Miltenyi Biotec-tested panel 13 (MBTP 13)

This application protocol describes the flow cytometric analysis of monocytes after spleen dissociation from healthy C57BL/6 mice. Mouse spleens are easily dissociated using the gentleMACS™ Dissociators to quickly obtain viable single cell suspensions ready for flow cytometric analysis.

Protocol

Gating strategy showing the analysis of monocytes from mouse spleen. Splenocytes from C57BL/6 mice were stained using the previously described panel. Samples were initially gated on single cells using FSC-H/FSC-A gating and on live cells using Viobility-negative gating (data not shown). CD45+ monocytes (A) lack expression of F4/80 (B) as well as T, B, and NK cell lineage markers (CD3, CD19, CD335; C), while expressing high levels of CD11b and Ly6C (C, D). In addition, analysis of the expression of the chemokine receptor CD192 helps to identify monocytes within mouse spleen (D).

Materials

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