Multi - color Flow Cytometry Mini Panel for Macrophage Identification

10 Jun 2025 by AntBio

Macrophages, tissue - resident white blood cells derived from monocytes (which originate from bone marrow precursor cells), are phagocytes. In vertebrates, they participate in both innate (non - specific) and cell - mediated (specific) immunity. Their main role is to phagocytose (engulf and digest) dead cells, cellular debris, and pathogens as fixed or free - ranging cells, and to activate lymphocytes or other immune cells, accelerating the immune response to pathogens.

This protocol offers a general method for total macrophage detection. To distinguish between monocyte - derived and tissue - resident macrophages, extra markers are needed.

Panel Recommendations

Marker	Fluorochrome	Max EX/EM (nm)	Corresponding Laser	Filter	Product Number	Format (1 T represents one test)
CD45	APC - Cy7	659/782	Red Laser	780/60	S0B1692	20/50/100/500 T available
Live/Dead Stain	FVS510	405/510	Violet Laser	450/50	abs47038974 - 200T	200 T
CD11b	PerCP - Cy5.5	482/676	Blue Laser	695/40	S0B5160	20/50/100/500 T available
F4/80	PE	496, 566/576	Blue or Yellow - Green Laser	575/26	abs1850269	25/100 μg available
CD86	Alexa Fluor® 488	494/517	Blue Laser	530/30	S0B1673	20/50/100/500 T available
CD206 (no membrane permeabilization required)	Alexa Fluor® 647	653/669	Red Laser	660/20	568808 568809	100 μg 25 μg
LY - 6G (optional)	Alexa Fluor® 700	697/717	Red Laser	730/45	S0B1700	20/50/100/500 T available

Notes:

Markers for M1 and M2 macrophages, such as CD80 and CD163, can be used as alternatives.

Since macrophage detection typically involves tissue samples, gating based on CD45 for immune cells and live/dead staining to exclude dead cells are essential.

F4/80 can show high false - positivity for neutrophils in tissues. Thus, adding LY - 6G is recommended to minimize neutrophil - related interference.

Detailed Explanation of Each Antibody Marker's Function:

CD antigen	Cellular expression	Molecular weight (kDa)	Functions
CD45	All hematopoietic cells	180–240 (multiple isoforms)	Tyrosine phosphatase, augments signaling through antigen receptor of B and T cells, multiple isoforms result from alternative splicing (see below)
CD11b	Myeloid and NK cells	170	αM subunit of integrin CR3 (associated with CD18): binds CD54, complement component iC3b, and extracellular matrix proteins
F4/80	macrophage	~160-180	Mediate cell adhesion and phagocytosis，articipate in antigen presentation，regulating tissue homeostasis and inflammatory response
CD86	Monocytes, activated B cells, dendritic cells	80	Ligand for CD28 and CTLA4
CD206	Macrophages, endothelial cells	175-190	Type I membrane glycoprotein; only known example of a C-type lectin that contains multiple C-type CRDs (carbohydrate-recognition domains); it binds high-mannose structures on the surface of potentially pathogenic viruses, bacteria, and fungi
LY-6G	Neutrophils (highly expressed), some inflammatory monocytes	25	Promote migration to the site of inflammation，regulate respiratory burst (ROS production), mediating NETosis (neutrophil extracellular trap)