Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Hamster

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, Simple Western, CyTOF-ready

Cited:

Immunohistochemistry-Paraffin, Western Blot, Bioassay, Mass Spectrometry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all EN-RAGE/S100A12 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human EN-RAGE/S100A12
Met1-Glu92
Accession # P80511

Specificity

Detects human EN-RAGE in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human EN‑RAGE/S100A12 Antibody

Detection of Human EN-RAGE/S100A12 antibody by Western Blot.

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 (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 antibody in Human Tonsil by Immunohistochemistry (IHC-P).

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; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human EN‑RAGE/S100A12 by Simple WesternTM.

Simple Western lane view shows lysates of human tonsil, loaded at 0.2 mg/mL. A specific band was detected for EN‑RAGE/S100A12 at approximately 11 kDa (as indicated) using 10 µg/mL of Goat Anti-Human EN‑RAGE/S100A12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1052). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of EN‑RAGE/S100A12 in Human peripheral blood monocytes by Flow Cytometry

Human peripheral blood monocytes were stained with Goat Anti-Human EN‑RAGE/S100A12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1052, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

Applications for Human EN‑RAGE/S100A12 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood monocytes

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Simple Western

10 µg/mL
Sample: Human tonsil

Western Blot

1 µg/mL
Sample: Human granulocytes

Reviewed Applications

Read 4 reviews rated 4.3 using AF1052 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: EN-RAGE/S100A12

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).

References

  1. Santamaria-Kisiel, L. et al. (2006) Biochem. J. 396:201.
  2. Leclerc, E. et al. (2009) Biochim. Biophys. Acta 1793:993.
  3. Wicki, R. et al. (1996) Cell Calcium 20:459.
  4. Moroz, O.V. et al. (2009) BMC Biochem. 10:11.
  5. Miranda, L.P. et al. (2001) FEBS Lett. 488:85.
  6. Vogl, T. et al. (1999) J. Biol. Chem. 274:25291.
  7. Xie, J. et al. (2007) J. Biol. Chem. 282:4218.
  8. Goyette, J. et al. (2009) J. Immunol. 183:593.
  9. Hatakeyama, T. et al. (2004) Eur. J. Biochem. 271:3765.
  10. Hofmann, M.A. et al. (1999) Cell 97:889.
  11. Yang, Z. et al. (2001) J. Leukoc. Biol. 69:986.
  12. Rouleau, P. et al. (2003) Clin. Immunol. 107:46.
  13. Yan, W.X. et al. (2008) J. Biol. Chem. 283:13035.
  14. Yang, Z. et al. (2007) J. Allergy Clin. Immunol. 119:106.
  15. Mikkelsen, S.E. et al. (2001) J. Neurochem. 79:767.
  16. Fuellen, G. et al. (2004) OMICS 8:334.

Long Name

Extracellular Newly Identified RAGE-binding Protein

Alternate Names

CAAF1, CAAFI, CAGC, CAGCS100, Calgranulin C, ENRAGE, MRP6, S100A12

Entrez Gene IDs

6283 (Human)

Gene Symbol

S100A12

UniProt

P80511

Additional EN-RAGE/S100A12 Products

Product Documents for Human EN‑RAGE/S100A12 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human EN‑RAGE/S100A12 Antibody

For research use only

Citations for Human EN‑RAGE/S100A12 Antibody

Customer Reviews for Human EN‑RAGE/S100A12 Antibody (4)

4.3 out of 5
4 Customer Ratings
5 Stars
25%
4 Stars
75%
3 Stars
0%
2 Stars
0%
1 Stars
0%

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Customer Images

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  • Human EN-RAGE/S100A12 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Recombinant protein
    Species: Human
    Verified Customer | Posted 09/29/2023
    Human EN‑RAGE/S100A12 Antibody AF1052
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 19944411
    Species: Human
    Verified Customer | Posted 01/05/2015
  • Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: See PMID 22609404
    Species: Human
    Verified Customer | Posted 01/05/2015
  • Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: See PMID 22609404
    Species: Other
    Verified Customer | Posted 01/05/2015

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Protocols

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