Protocol for assessing anti-Siglec-9 antibody-mediated tumor phagocytosis by macrophages using pH-sensitive dyes and flow cytometry

STAR Protocols 2026 Open access

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Authors: Saitong Muneekaew, Thunchanok Nuchphongsai, Pasut Sasithong, Kritayaporn Saiprayong, Kamonphan Rungrueang, Methichit Wattanapanitch
DOI: 10.1016/j.xpro.2026.104590
Cited by: 1
References: 7

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 + 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. 1

Fields of Study & Subjects

Phagocytosis and Immune Regulation Monoclonal and Polyclonal Antibodies Research Immune cells in cancer

Publication details

Journal / source
STAR Protocols
Publisher
Elsevier BV
Volume
7
Issue
2
Pages
104590–104590
Publication date
20 May 2026
Language
EN