Detection of Human, Mouse, and Rat RAGE by Western Blot. Western blot shows lysates of human, mouse, and rat lung tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat RAGE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1145) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for RAGE at approximately |
45-55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
|Detection of Human RAGE by Western Blot. Western blot shows lysates of human lung tissue and HEK001 human epidermal keratinocyte cell line. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat RAGE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1145) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for RAGE at approximately 43 kDa under non-reducing (NR) conditions and 50 kDa under reducing (R) conditions (as indicated). This experiment was conducted using Immunoblot Buffer Group 5.|
|RAGE in Human Alzheimer's Disease Brain. RAGE was detected in immersion fixed paraffin-embedded sections of human Alzheimer's disease brain (cerebellum) using 15 µg/mL Goat Anti-Human/Mouse/Rat RAGE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1145) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
|RAGE in Human Lung. RAGE was detected in immersion fixed paraffin-embedded sections of human lung using Goat Anti-Human/Mouse/Rat RAGE Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1145) at 0.5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membranes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.|
Advanced glycation endproducts (AGE) are adducts formed by the non-enzymatic glycation or oxidation of macromolecules. AGE forms during aging and its formation is accelerated under pathophysiologic states such as diabetes, Alzheimer’s disease, renal failure and immune/inflammatory disorders. Receptor for Advanced Glycation Endoproducts (RAGE), named for its ability to bind AGE, is a multi-ligand receptor belonging the immunoglobulin (Ig) superfamily. Besides AGE, RAGE binds amyloid beta -peptide, S100/calgranulin family proteins, high mobility group B1 (HMGB1, also know as amphoterin) and leukocyte integrins.
The human RAGE gene encodes a 404 amino acid residues (aa) type I transmembrane glycoprotein with a 22 aa signal peptide, a 320 aa extracellular domain containing an Ig-like V-type domain and two Ig-like Ce-type domains, a 21 aa transmembrane domain and a 41 aa cytoplasmic domain. The V-type domain and the cytoplasmic domain are important for ligand binding and for intracellular signaling, respectively. Two alternative splice variants, lacking the V-type domain or the cytoplasmic tail, are known. RAGE is highly expressed in the embryonic central nervous system. In adult tissues, RAGE is expressed at low levels in multiple tissues including endothelial and smooth muscle cells, mononuclear phagocytes, pericytes, microglia, neurons, cardiac myocytes, and hepatocytes. The expression of RAGE is upregulated upon ligand interaction. Depending on the cellular context and interacting ligand, RAGE activation can trigger differential signaling pathways that affect divergent pathways of gene expression. RAGE activation modulates varied essential cellular responses (including inflammation, immunity, proliferation, cellular adhesion, and migration) that contribute to cellular dysfunction associated with chronic diseases such as diabetes, cancer, amyloidoses, and immune or inflammatory disorders.
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