Mouse CCL17/TARC Antibody

Chemotaxis Induced by CCL17/TARC and Neutralization by Mouse CCL17/TARC Antibody.

Recombinant Mouse CCL17/ TARC (Catalog # 529-TR) chemoattracts the BaF3 mouse pro-B cell line transfected with human CCR4 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Mouse CCL17/TARC (0.02 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse CCL17/TARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF529). The ND₅₀ is typically 0.2-0.6 µg/mL.

Detection of Mouse CCL17/TARC by Immunocytochemistry/ Immunofluorescence

Cd248 disruption reduced infiltration and pro-fibrotic phenotype switching of kidney macrophages during renal fibrosis. (a) Representative images of F4/80 immunostaining for macrophages in control and D14 UUO kidneys. F4/80+ staining was brown. Original magnification, × 100. Scale bar, 100 μm. (b) Quantification of F4/80+ areas on LPF images of kidney sections taken at 100 × magnification n = 10. (c). Representative images of green fluorescent protein (GFP)+ circulation-derived cells and F4/80+ (red) macrophages in control and day 3 (D3) UUO-kidneys of WT and Cd248–/– parabionts joined surgically to transgenic GFP (GFPTg) mice. Arrowhead indicates an F4/80+GFP+ macrophage. Original magnification, × 100. Scale bar, 100 μm. (d) Quantification of GFP+, F4/80+ and F4/80+GFP+ cells on LPF images of control (upper panel) and D3 UUO (lower panel) kidneys of WT and Cd248–/– parabionts. n = 3. (e) qPCR of genes encoding cytokines and enzymes in macrophages isolated from D7 UUO kidneys. n = 4. (f) qPCR of Nos2, Arg1 and Ccl17 in lipopolysaccharide (LPS) and interferon gamma (IFN gamma)–primed RAW264.7 macrophages co-cultured with D7 UUO-kidney myofibroblasts isolated from WT or Cd248–/– mice in the Transwell system. RAW264.7 macrophages co-cultured with medium were used as control. n = 4. (g) qPCR of Ccl17 in WT bone marrow–derived macrophages (BMDMs, Mϕ) co-cultured with medium only (control) or with WT or Cd248–/– UUO-kidney myofibroblasts (MF) in the same dish. Recombinant CD248 (rCD248) was included in the culture as indicated. n = 5. Data are expressed as means ± standard errors of the mean. *P < 0.05, **P < 0.01, ***P < 0.001 by one-way ANOVA with post hoc Tukey’s multiple comparisons test in (b,g) and unpaired t-test in (d,e,f). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33033277), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CCL17/TARC by Immunocytochemistry/ Immunofluorescence

Cd248 disruption reduced infiltration and pro-fibrotic phenotype switching of kidney macrophages during renal fibrosis. (a) Representative images of F4/80 immunostaining for macrophages in control and D14 UUO kidneys. F4/80+ staining was brown. Original magnification, × 100. Scale bar, 100 μm. (b) Quantification of F4/80+ areas on LPF images of kidney sections taken at 100 × magnification n = 10. (c). Representative images of green fluorescent protein (GFP)+ circulation-derived cells and F4/80+ (red) macrophages in control and day 3 (D3) UUO-kidneys of WT and Cd248–/– parabionts joined surgically to transgenic GFP (GFPTg) mice. Arrowhead indicates an F4/80+GFP+ macrophage. Original magnification, × 100. Scale bar, 100 μm. (d) Quantification of GFP+, F4/80+ and F4/80+GFP+ cells on LPF images of control (upper panel) and D3 UUO (lower panel) kidneys of WT and Cd248–/– parabionts. n = 3. (e) qPCR of genes encoding cytokines and enzymes in macrophages isolated from D7 UUO kidneys. n = 4. (f) qPCR of Nos2, Arg1 and Ccl17 in lipopolysaccharide (LPS) and interferon gamma (IFN gamma)–primed RAW264.7 macrophages co-cultured with D7 UUO-kidney myofibroblasts isolated from WT or Cd248–/– mice in the Transwell system. RAW264.7 macrophages co-cultured with medium were used as control. n = 4. (g) qPCR of Ccl17 in WT bone marrow–derived macrophages (BMDMs, Mϕ) co-cultured with medium only (control) or with WT or Cd248–/– UUO-kidney myofibroblasts (MF) in the same dish. Recombinant CD248 (rCD248) was included in the culture as indicated. n = 5. Data are expressed as means ± standard errors of the mean. *P < 0.05, **P < 0.01, ***P < 0.001 by one-way ANOVA with post hoc Tukey’s multiple comparisons test in (b,g) and unpaired t-test in (d,e,f). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33033277), licensed under a CC-BY license. Not internally tested by R&D Systems.