MBTP 16: Immunophenotyping of basophils from mouse spleen using flow cytometry

Miltenyi Biotec-tested panel 16 (MBTP 16)

This application protocol describes the flow cytometric analysis of basophils 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 basophils from mouse spleen. Splenocytes from C57BL/6 mice were stained using the previously described panel to identify basophils. 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+ basophils (A) express CD11b and lack expression of T, B, and NK cell lineage markers CD3, CD19, and CD335 (B). Within CD11b+ cells, basophils express CD49b while lacking expression of CD117 (C). Within CD49b+ cells, basophils can be efficiently identified by the co-expression of the high-affinity IgE receptor (FcεRIα) and CD200R3 (D). 

Materials

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