Intracellular Staining by Flow Cytometry
|Detection of CXCL17/VCC-1 in A549 Human Cell Line by Flow Cytometry. A549 human lung carcinoma cell line was stained with Mouse Anti-Human CXCL17/VCC-1 Monoclonal Antibody (Catalog # MAB4207, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.|
|CXCL17/VCC‑1 in A549 Human Cell Line. CXCL17/VCC‑1 was detected in immersion fixed A549 human lung carcinoma cell line using Mouse Anti-Human CXCL17/VCC‑1 Monoclonal Antibody (Catalog # MAB4207) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI(blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
CXCL17, also known as dendritic cell and monocyte chemokine-like protein (DMC) and VEGF-correlated chemokine-1 (VCC-1), is a secreted molecule with a size and predicted three-dimensional folding pattern similar to that of chemokines CXCL8/IL-8 and CXCL14/BRAK (1, 2). It has no predicted N-glycosylation site. Cleavage of a 23 amino acid (aa) signal sequence yields the mature 96 aa human CXCL17. CXCL17 is constitutively produced by airway and intestinal epithelium (1). It induces the chemotaxis of quiescent, but not LPS-activated peripheral blood monocytes and dendritic cells (1). CXCL17 expression is increased in endothelial cells when they are induced to form tubes in vitro (2). Transgenic overexpression in NIH3T3 cells causes upregulation of proteins such as VEGF and FGF basic, and increases cell growth rate and tumorigenicity (2). CXCL17, plus two other chemokines that play roles in angiogenesis, CXCL1/GRO and CXCL8/IL-8, show a correlated expression pattern with VEGF in primary lung, breast and esophageal tumors (2). CXCL17 is, therefore, suggested to play a role in tumor angiogenesis. Mature human CXCL17 shares 73%, 71% and 64% amino acid sequence identity with bovine, mouse and rat CXCL17, respectively.
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