Human/Mouse/Rat RAGE Antibody

Detection of Human RAGE by Western Blot. Western blot shows lysates of human lung tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat RAGE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1179) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for RAGE at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse and Rat RAGE by Western Blot. Western blot shows lysates of mouse lung tissue and rat lung tissue. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human/Mouse/Rat RAGE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1179) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for RAGE at approximately 40-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

RAGE in Mouse Brain. RAGE was detected in immersion fixed frozen sections of mouse brain using Goat Anti-Mouse/Rat RAGE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1179) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to neurons. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Detection of Mouse AGER by Immunohistochemistry RAGE contributes to maintenance of pulmonary mechanics and structure.Quantification of mean chord length (A) were performed by stereological analysis of alveolar parenchyma; n ≥ 4 per group. (B) Representative histology (hematoxylin and eosin staining) of ten months old WT and RAGE-/- mice; scale bars: 200μm. Respiratory system compliance (C) and respiratory system elastance (D) were determined in two, four and ten months old WT and RAGE-/- mice using invasive pulmonary function measurements; n ≥ 7 per group. (E) Concentration of serum protein albumin in BALF of two months old mice; n = 10. Data are shown as mean ± SEM. *p < 0.05 and **p < 0.01. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0180092), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse AGER by Immunohistochemistry RAGE is expressed on alveolar epithelial cells and alveolar macrophages.(A) Immunostaining for RAGE protein in WT and RAGE-/- lung tissue. (B) Isolated alveolar epithelial cells (AEC) (n = 7 per group) and alveolar macrophages (AM) (n = 4 per group) were analyzed for RAGE mRNA expression using qRT-PCR. Data are shown as mean ± SEM. Scale bar: 100μm. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0180092), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse AGER by Immunohistochemistry RAGE is expressed on alveolar epithelial cells and alveolar macrophages.(A) Immunostaining for RAGE protein in WT and RAGE-/- lung tissue. (B) Isolated alveolar epithelial cells (AEC) (n = 7 per group) and alveolar macrophages (AM) (n = 4 per group) were analyzed for RAGE mRNA expression using qRT-PCR. Data are shown as mean ± SEM. Scale bar: 100μm. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0180092), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Human/Mouse/Rat RAGE Antibody by Western Blot Metformin activates AMPK and inhibits AGEs-induced RAGE expression and NF kappa B activation. (a) BMDMs were divided into 4 groups: control, AGEs, MET, and AGEs + MET group. In AGEs group, cells were cultured with AGEs at 200 mg/L for 24 h; in MET group, cells were cultured with metformin at 2.0 μM for 24 h; in AGEs + MET group, cells were pretreated with metformin for 60 min and then cultured with AGEs at 200 mg/L for 24 h; in control group, cells were cultured with BSA at 200 mg/L for 24 h. Western blot analysis was performed to measure protein levels of RAGE and phosphorylated AMPK (p-AMPK). Tubulin was used as internal control. (b) BMDMs were pretreated with or without metformin (2.0 μM) for 60 min before AGEs (200 mg/L) stimulation for different time intervals (0, 30, 60, and 180 min). Protein levels of NF kappa B-p65 (p65) and phosphorylated NF kappa B-p65 (p-p65) were measured by western blot. Tubulin was used as internal control. Bar graphs represent the results (mean ± SD) of three independent experiments. One-way ANOVA was applied and all the overall ANOVA was significant. #p > 0.05; ∗p < 0.05; ∗∗p < 0.01; and ∗∗∗p < 0.001 when compared between selected groups. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27761470), licensed under a CC-BY license. Not internally tested by R&D Systems.