MBTP 17: Immunophenotyping of dendritic cells from mouse spleen using flow cytometry

Miltenyi Biotec-tested panel 17 (MBTP 17)

This application protocol describes the flow cytometric analysis of dendritic cells (DCs) 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 dendritic cell (DCs) populations from healthy mouse spleen. Splenocytes from C57BL/6 mice were stained using the previously described DC 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). Conventional (CD11c+ MHC class II+) and plasmacytoid (Siglec-H+ CD317+) DCs were identified (A, D). Two major conventional DC (cDC) subsets, cDC1 and cDC2, can be found in spleen. The cDC1 subset is described as cross-presenting DC type and characteristically expresses XCR1 and lacks expression of CD172a. Non-cross presenting DCs (cDC2) express the opposite phenotype (XCR1 CD172a+) (B). Alternatively, cDC1 and cDC2 subsets can be differentiated using XCR1 and CD11b (C).

Materials

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