Human/Mouse Arginase 1/ARG1 APC-conjugated Antibody

Detection of Arginase 1/ARG1 in HepG2 Human Cell Line by Flow Cytometry. HepG2 human hepatocellular carcinoma cell line was stained with Sheep Anti-Human/Mouse Arginase 1/ARG1 APC-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # IC5868A, filled histogram) or isotype control antibody (Catalog # IC016A, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Detection of Arginase 1/ARG1 in Hepa 1-6 Mouse Cell Line by Flow Cytometry. Hepa 1-6 mouse hepatoma cell line was stained with Sheep Anti-Human/Mouse Arginase 1/ARG1 APC-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # IC5868A, filled histogram) or isotype control antibody (Catalog # IC016A, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Detection of Mouse Arginase 1/ARG1/liver Arginase by Flow Cytometry ILC2 transfers to apoE−/− mice increase peritoneal B1 cells, eosinophils and AAMs. Flow cytometry gating strategies of B1 cells identified as CD19+B220lowCD11b+IgM+ (a), eosinophils as CD45+SiglecF+CD11b+ cells (d) and AAMs as CD45+CD11b+F480+Arg1+ cells (i) of apoE−/− mice for both control and ILC2 groups respectively as indicated. Percentages and numbers of peritoneal B1 cells (b, c), eosinophils (e, f) and AAMs (j, k). CD11c+ expression from gated eosinophils and respective cell numbers of activated eosinophils identified as CD45+SiglecF+CD11b+CD11c+cells (g, h). Mann Whitney U test, **P < 0.01, ***P < 0.001, ****P < 0.0001. Each data point represents one mouse Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31823769), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Arginase 1/ARG1/liver Arginase by Flow Cytometry Flow cytometry and qRT/PCR analyses of immune cell infiltrates. (A) Flow cytometry gating strategy for macrophages (CD11b+CD45hiLy6G-), microglia (CD11b+CD45loLy6G-), and neutrophils (CD11b+CD45hiLy6G+). (B) Macrophages or microglia were further characterized as alternatively activated (M2) CD206+Ym1+Arg1+ or (C) classically activated (M1) CD86+MHCII+iNOS+. (D) Flow cytometry gating for T cell subsets: Th1 = CD4+IFN-gamma +, Th17 = CD4+IL17+, Treg = CD4+CD25+FOXP3. (E, F, G) Flow cytometry of cells isolated from pooled brain and spinal cord showed that the only significant difference in total numbers of T cell subtypes isolated from the CNS at 21 dpi was a decrease in the ratio of IFN-gamma + to IL-17+ cells in the astroglial CXCL10 knockout mice (n = 3 mice/group, P = 0.0063). (H, I) No significant differences between astroglial CXCL10 knockout and control mice in total numbers of macrophages, microglia, and neutrophils (H) or M1 and M2 subtypes of macrophages and microglia (I) isolated from the CNS at 21 dpi (n = 3). (J) qRT/PCR of spinal cord tissue isolated at 14 dpi normalized to the housekeeping gene HSP90; CXCR3 expression was significantly upregulated in astroglial CXCL10 knockout mice compared to controls (P = 0.0157). IFN-gamma, FOXP3, ROR gamma t, iNOS, and arginase-1 mRNA levels were not significantly different between astroglial CXCL10 knockout and control mice at 14 dpi (n = 6 mice/group). Vertical bars = SEMs. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24924222), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Arginase 1/ARG1/liver Arginase by Flow Cytometry Tumor permissive microenvironment in Brca1 MT mammary glands.aS100a9/S100a8 mRNA expression in the subpopulations of luminal and stromal cells of WT 4-month mammary gland (WTV4MG) and MT 4-months-old virgin mammary gland (MTV4MG) (n = 3 mice). b Protein level of S100a9 in both WT (B477) and MT (G600) mammary epithelial cell lines and tumor tissues by Western blots (n = 3 individual experiment-up and n = 3 mice-down). c Co-staining of S100a9 (red) and CK18 (green) with antibodies on WTV4MG, MTV4MG, WTV6MG, and MTV6MG tissues (n = 3 pairs in each group, Scale bar: 20 μM). d The S100a9 and Arg1 positive cell populations by FACS analysis from the blood and mammary tissues of both WT and MT mice at 4-month and 6-month, respectively (FACS gating strategies see in Supplementary Fig. 8c, n = 3 mice/ group). e Co-staining with S100a9 (red) and CD206 (green) antibodies (left panel) and co-staining with S100a9 (red) and CK18 (green) antibodies (right panel) on tumor-adjacent tissues by IF (40X confocal microscope, Scale bar: 20 μM.) (n = 3 mice and 3 individual experiment). f Secreted S100a9 proteins (left) from both tumor cell and MDSC cells in tumor-adjacent mammary gland (n = 3 mice) and present in the supernatant of cultured cancer cells (right) (n = 3 individual experiment, Scale bar: 10 μM). g Protein levels of S100a9, TGF-beta, and Il-10 in mammary gland tissues of both WT and Brca1 MT mice at 4-month (n = 3 mice). h Protein levels of S100a9, TGF-beta, and IL-10 in mammary tissues of both WT and Brca1 MT mice at 6-month (n = 3 mice). The data are expressed as means ± SD (a) and P values determined by unpaired two-tailed Student’s t test. The experiments were independently repeated three times with similar results (a, b). Source data are provided as a Source data file. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35304461), licensed under a CC-BY license. Not internally tested by R&D Systems.