APC Anti-Mouse CD163 Monoclonal Antibody for Flow Cytometry - Q2VLH6

Core Product Specifications and Parameters

Biological Background: CD163 Function and Localization

• CD163, also known as scavenger receptor cysteine-rich type 1 protein M130, is a member of the scavenger receptor cysteine-rich (SRCR) superfamily. It is expressed primarily on monocytes and mature macrophages, including Kupffer cells in the liver, red pulp macrophages in the spleen, and mesenteric lymph node macrophages.
• The protein functions in the clearance and endocytosis of hemoglobin/haptoglobin (Hb/Hp) complexes by macrophages. This process protects tissues from oxidative damage caused by free hemoglobin and facilitates iron recycling via heme breakdown.
• CD163 binding to Hb/Hp complexes is calcium-dependent and pH-dependent. Upon ligand engagement, CD163 triggers intracellular signaling cascades involving tyrosine kinase-dependent calcium mobilization and inositol triphosphate production, leading to secretion of IL-6 and CSF-1.
• A soluble form (sCD163) is generated by ectodomain shedding. Elevated levels of sCD163 are associated with inflammatory conditions, where it may play an anti-inflammatory role by acting as a decoy receptor.
• Structurally, CD163 is a single-pass type I transmembrane protein with a signal peptide, multiple SRCR domains, and a short cytoplasmic tail. It is heavily glycosylated and contains disulfide bonds, contributing to its stability and ligand-binding properties.
• The human CD163 gene is located at chromosome 12p13.31, while the mouse ortholog is on chromosome 6. The protein undergoes alternative splicing, producing variants with potential functional differences.
• CD163 is recognized as an acute phase response marker; its expression is upregulated by anti-inflammatory stimuli (e.g., glucocorticoids, IL-10) and downregulated by pro-inflammatory signals (e.g., LPS, TNF-α).

Experimental Guidance and Technical Tips

• For flow cytometry, use the directly conjugated APC antibody. Perform titration to determine the optimal working concentration for your specific cell type and experimental conditions, and always include an appropriate isotype control.
• This antibody targets amino acids 39–260 of the extracellular domain, which is accessible on the cell surface. It is suitable for live-cell staining without permeabilization.
• Validate reactivity in relevant mouse tissue samples known to express CD163, such as spleen, liver, or lymph nodes. Enzymatic digestion may improve macrophage recovery for staining.
• Because CD163 expression can be modulated, consider treating cells with IL-10 or glucocorticoids to enhance detection in vitro if expression levels are low.
• Use Fc receptor blocking (e.g., anti-CD16/CD32) prior to staining to minimize non-specific binding to Fc receptors on macrophages.

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