Mouse RAGE/AGER Antibody

Detection of Mouse and Rat RAGE by Western Blot.

Western blot shows lysates of mouse lung tissue, Neuro-2A mouse neuroblastoma cell line, and rat lung tissue. PVDF membrane was probed with 1 µg/mL of Rat Anti-Mouse RAGE Monoclonal Antibody (Catalog # MAB11795) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (HAF005). Specific bands were detected for RAGE at approximately 45 to 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of RAGE in Mouse Splenocytes by Flow Cytometry.

Mouse splenocytes stimulated to induce Th1 cells were stained with Rat Anti-Mouse CD4 PE-conjugated Monoclonal Antibody (FAB554P) and either (A) Rat Anti-Mouse RAGE Monoclonal Antibody (Catalog # MAB11795) or (B) Rat IgG_2AIsotype Control (MAB006) followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (F0113). View our protocol for Staining Membrane-associated Proteins.

Detection of Mouse AGER by Immunocytochemistry/Immunofluorescence

Treatment with anti-M7 prevents inflammatory leukocyte recruitment in vitro and in vivo. a Murine peritoneal macrophages were incubated on dishes coated with mouse CD40L in the presence of 10μg/ml of IgG, anti-M7, or anti-Mac-1 (clone M1/70). Adhering cells were normalized to % of the IgG-treatment. b Murine RAW-cell adhesion on TNF alpha-primed murine endothelial cells under physiological flow in the presence of IgG or anti-M7 antibodies (10 μg/ml). c–f C57Bl/6 mice were injected i.p. with 200 ng TNF alpha and 15 min before microscopy with Fab fragments of an IgG control antibody, or antibodies against RAGE, ICAM-1, LFA-1, Mac-1 (clone M1/70), or anti-M7 (100 μg i.p.). Leukocyte recruitment was monitored by intravital microscopy 4 h later: Adhering (d) and rolling leukocytes (e) were quantified as % of IgG. (f) Cumulative frequency of leukocyte rolling velocity. g Leukocyte adhesion in intravital microscopy in IgG or anti-M7 treated WT and Mac-1−/− mice. h, i Number of CX3CR1-GFP+ monocytes (white arrows) in the para-vascular space 4 h after treatment with IgG or anti-M7 and 200ng TNF alpha. j Confocal in vivo imaging of the greater omentum after local stimulation with TNF alpha and i.v. injection of the indicated antibodies (100 μg) in LysM-GFP reporter mice. LysM-GFP+ myeloid cells were tracked over time. Time lapse and cell tracking is shown over 15 min (right panel). k Plasma cytokine levels of mice subjected to intravital microscopy after IgG or anti-M7 Fab treatment. l Peritoneal exudate cells (PECs) 72 h after induction of a sterile peritonitis and the indicated antibody treatment. Scale bars represent 100μm (c, h, j). Error bars indicate mean ± SEM. Statistical significance was assessed by a two-sided, unpaired Student’s T-test between the indicated groups (a, b, g, i, k, l) or in comparison to IgG-treatment (d, e). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. N ≥ 3 independent experiments (a, b). N ≥ 10 mice per group (d, e, f, g), N ≥ 6 mice per group (l, h) Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29410422), licensed under a CC-BY license. Not internally tested by R&D Systems.