Protocol for assessing anti-Siglec-9 antibody-mediated tumor phagocytosis by macrophages using pH-sensitive dyes and flow cytometry
1
Issued Date
2026-06-19
Resource Type
eISSN
26661667
Scopus ID
2-s2.0-105039528008
Journal Title
STAR Protocols
Volume
7
Issue
2
Rights Holder(s)
SCOPUS
Bibliographic Citation
STAR Protocols Vol.7 No.2 (2026)
Suggested Citation
Muneekaew S., Nuchphongsai T., Sasithong P., Saiprayong K., Rungrueang K., Wattanapanitch M. Protocol for assessing anti-Siglec-9 antibody-mediated tumor phagocytosis by macrophages using pH-sensitive dyes and flow cytometry. STAR Protocols Vol.7 No.2 (2026). doi:10.1016/j.xpro.2026.104590 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/116959
Title
Protocol for assessing anti-Siglec-9 antibody-mediated tumor phagocytosis by macrophages using pH-sensitive dyes and flow cytometry
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Abstract
Here, we present a protocol to assess and quantify macrophage-mediated cancer cell phagocytosis using pH-sensitive dyes and flow cytometry. We describe steps for isolating and sorting CD14<sup>+</sup> monocytes from leukocyte reduction system (LRS) cones, differentiating monocyte-derived macrophages, and preparing K562 leukemic cells for co-culture. We then detail procedures for quantifying phagocytic activity using flow cytometry and pH-sensitive dyes and visualizing internalized K562 cells using a fluorescence microscope. This technique enables the study of macrophage phagocytic function and supports application in cancer immunotherapy research. For complete details on the use and execution of this protocol, please refer to Muneekaew et al. <sup>1</sup>