Detection of Human EN‑RAGE/S100A12 by Western Blot. Western blot shows lysate of human granulocytes. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human EN‑RAGE/S100A12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1052) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EN‑RAGE/S100A12 at approximately 10 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
EN‑RAGE/S100A12 in Human Tonsil. EN‑RAGE/S100A12 was detected in immersion fixed paraffin-embedded sections of human tonsil using 15 µg/mL Goat Anti-Human EN‑RAGE/S100A12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1052) 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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
EN-RAGE, also known as S100A12 and Calgranulin C, is a 10 kDa member of the S100 (soluble in 100% saturated ammonium sulfate) family of EF-hand calcium-binding proteins. Like other S100 proteins, S100A12 is small and generally acidic (1-3). EN-RAGE forms noncovalent homodimers in the absence of divalent cations (4-6); calcium and zinc promote the formation of higher order assemblies including tetramers and hexamers (4, 7, 8). The ability of S100A12 to chelate zinc enables it to inhibit the zinc-dependent metalloproteases MMP-2, -3, and -9 (8). S100A12 also forms heterodimers with S100A9 and binds to RAGE (Receptor for Advanced Glycation End-products), Annexin V, and several cytosolic enzymes involved in energy metabolism (9, 10). The hexameric form of EN-RAGE in particular binds RAGE with high affinity (7). EN-RAGE induces a variety of inflammatory responses including the in vivo recruitment of neutrophils, monocytes, and mast cells and the activation of mast cells and vascular endothelial cells (9, 11-14). EN-RAGE is found at elevated levels under inflammatory conditions such as asthma, gout, rheumatoid arthritis synovial fluid, and atherosclerosis (8, 12, 14). S100A12 also promotes neurite outgrowth in isolated hippocampal neurons (15). An ortholog of S100A12 has not been identified in rodents, but the human protein is functional in mice and rats (11-16).
Santamaria-Kisiel, L. et al. (2006) Biochem. J. 396:201.
Leclerc, E. et al. (2009) Biochim. Biophys. Acta 1793:993.
Wicki, R. et al. (1996) Cell Calcium 20:459.
Moroz, O.V. et al. (2009) BMC Biochem. 10:11.
Miranda, L.P. et al. (2001) FEBS Lett. 488:85.
Vogl, T. et al. (1999) J. Biol. Chem. 274:25291.
Xie, J. et al. (2007) J. Biol. Chem. 282:4218.
Goyette, J. et al. (2009) J. Immunol. 183:593.
Hatakeyama, T. et al. (2004) Eur. J. Biochem. 271:3765.
Hofmann, M.A. et al. (1999) Cell 97:889.
Yang, Z. et al. (2001) J. Leukoc. Biol. 69:986.
Rouleau, P. et al. (2003) Clin. Immunol. 107:46.
Yan, W.X. et al. (2008) J. Biol. Chem. 283:13035.
Yang, Z. et al. (2007) J. Allergy Clin. Immunol. 119:106.
Mikkelsen, S.E. et al. (2001) J. Neurochem. 79:767.
Fuellen, G. et al. (2004) OMICS 8:334.
Extracellular Newly Identified RAGE-binding Protein
Entrez Gene IDs:
CAAF1; CAAF1Neutrophil S100 protein; CAAFI; CAGC; CAGCS100 calcium binding protein A12 (calgranulin C); CAGCS100; Calcium-binding protein in amniotic fluid 1; Calgranulin C; calgranulin-C; CGRPEN-RAGE; ENRAGE; Extracellular newly identified RAGE-binding protein; MRP6; p6calgranulin C; protein S100-A12; S100 calcium binding protein A12; S100 calcium-binding protein A12 (calgranulin C); S100 calcium-binding protein A12; S100A12
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The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
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