详细说明
Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse, and rat RAGE in Western blots. In direct ELISAs, less than 2% cross-reactivity with recombinant canine RAGE is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse RAGE
Gln24-Ala342
Accession # O35444Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated
Applications
Recommended
ConcentrationSample
Western Blot
0.1-1 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
Blockade of Receptor-ligand Interaction
In a functional ELISA, 15-35 µg/mL of this antibody will block 50% of the binding of 500 ng/mL of biotinylated AGE-BSA to immobilized Recombinant Mouse RAGE Fc Chimera (Catalog # ) coated at 5 µg/mL (100 µL/well). At 166 μg/mL, this antibody will block >90% of the binding.
Please Note: Optimal dilutions should be determined by each laboratory for each application. are available in the Technical Information section on our website.
Data Examples
Western Blot | 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 # ). A specific band was detected for RAGE at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using . |
Western Blot | 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 # ). Specific bands were detected for RAGE at approximately 40-50 kDa (as indicated). This experiment was conducted under reducing conditions and using . |
Immunohistochemistry | 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 # ) and counterstained with DAPI (blue). Specific staining was localized to neurons. View our protocol for . |
Preparation and Storage
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: RAGE
Advanced glycation endproducts (AGE) are adducts formed by the non-enzymatic glycation or oxidation of macromolecules (1). 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 multiligand 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 (1, 2).
The mouse RAGE gene encodes a 403 amino acid (aa) residue type I transmembrane glycoprotein with a 22 aa signal peptide, a 319 aa extracellular domain containing a Ig-like V-type domain and two Ig-like Ce-type domains, a 21 aa transmembrane domain and a 41 aa cytoplasmic domain (3). 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 (1, 4). RAGE is highly expressed in the embryonic central nervous system (5). 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 (6). 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 (1, 7). 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 (1).
References:
Schmidt, A. et al. (2001) J. Clin. Invest. 108:949.
Chavakis, T. et al. (2003) J. Exp. Med. 198:507.
Renard, C. et al. (1997) Mol. Pharmacol. 52:54.
Yonekura, H. et al. (2003) Biochem. J. 370:1097.
Hori, O. et al. (1995) J. Biol. Chem. 270:25752.
Brett, J. et al. (1993) Am. J. Pathol. 143:1699.
Valencia, J.V. et al. (2004) Diabetes 53:743.
Long Name:
Receptor for Advanced Glycation End Products
Entrez Gene IDs:
177 (Human); 11596 (Mouse); 81722 (Rat)
Alternate Names:
AGER; EC 2.7.11.22; MOK protein kinase; MOKMAPK/MAK/MRK overlapping kinase; RAGE; RAGE-1; RAGE1renal cell carcinoma antigen (MOK protein kinase); Renal tumor antigen 1; renal tumor antigen