| Advanced glycation endproducts (AGEs) are adducts formed by the nonenzymatic glycation of macromolecules. AGE formation is accelerated in oxidative and hyperglycemic conditions, diabetes, renal failure, atherosclerosis, Alzheimers disease, arthritis, and in normal aging (1-5). Receptor for advanced glycation endproducts (RAGE) is a 35kD type I transmembrane protein belonging the immunoglobulin superfamily. Besides AGEs, RAGE binds b-amyloid peptide, S100/calgranulin family proteins, HMGB1/amphoterin, and leukocyte integrins (6-9). Mature canine RAGE consists of a 383 amino acid (aa) extracellular domain (ECD) with one Ig-like V-type domain and two Ig-like C-type domains, a 23 aa transmembrane segment, and a 43 aa cytoplasmic domain (10). Within the ECD, canine RAGE shares 73%-77% aa sequence identity with human, mouse, and rat RAGE. In human, soluble forms of RAGE are generated by alternate splicing and are associated with multiple disease states (11, 12). RAGE is expressed in the embryonic central nervous system and on macrophages, monocytes, smooth muscle cells, and endothelial cells (13-15). It is upregulated in response to AGE accumulation, and its activation induces a broad proinflammatory response (6, 15). The increased production of reactive oxygen species during inflammation promotes additional AGE formation and RAGE upregulation, a cycle that exacerbates diabetic complications and inflammation-induced tissue injury (2, 4). A DNA sequence encoding the extracellular domain of canine RAGE (Met 1-Val 339, Accession ABA18645) was fused to a six-histidine tag at the C-terminus in a mouse myeloma cell line, NS0. Calculated molecular mass of 34.6kD. The recombinant canine RAGE migrates as an ~40-44kD protein in SDS-PAGE under reducing conditions. Activity: Measured by its ability to bind biotinylated advanced glycation endproducts of bovine serum albumin (AGE-BSA) Storage and Stability: Lyophilized powder may be stored at 4C for short-term only. Reconstitute to nominal volume by adding sterile PBS and store at -20C. Reconstituted product is stable for 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
