Key Product Details

Species Reactivity

Validated:

Human

Applications

CyTOF-ready, ELISA, Immunohistochemistry, Intracellular Staining by Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 900027

View all S100A8/S100A9 Heterodimer Antibodies »

Product Summary for Human S100A8/S100A9 Heterodimer Antibody

Immunogen

E. coli-derived recombinant human S100A8/S100A9 heterodimer
Met1-Glu93 (S100A8) & Thr2-Pro114 (S100A9)
Accession # P05109 (S100A8) and P06702 (S100A9)

Specificity

Detects human S100A8/S100A9 heterodimer in direct ELISAs. In direct ELISAs, no detection of recombinant human S100A8 or S100A9 monomers was observered.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human S100A8/S100A9 Heterodimer Antibody

S100A8/S100A9 Heterodimer antibody in Human Colon Cancer Tissue by Immunohistochemistry (IHC-P).

S100A8/S100A9 Heterodimer in Human Colon Cancer Tissue.

S100A8/S100A9 Heterodimer was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using Mouse Anti-Human S100A8/S100A9 Heterodimer Monoclonal Antibody (Catalog # MAB45702) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to connective tissue fibroblasts. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of S100A8/A9 antibody in Human PBMCs antibody by Flow Cytometry.

Detection of S100A8/A9 in Human PBMC Monocytes by Flow Cytometry.

Human peripheral blood monocytes (PBMC) were stimulated with LPS (1 ug/mL) and Monensin overnight, then stained with either (A) Mouse Anti-Human S100A8/A9 Monoclonal Antibody (Catalog # MAB45702) or (B) Mouse IgG1Isotype Control (MAB002) followed by Goat anti-Mouse IgG APC-conjugated Secondary Antibody (F0101B) and Mouse Anti-Human CD14 PE-conjugated Monoclonal Antibody (FAB3832P). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer (FC005). View our protocol for Staining Intracellular Molecules.

Applications for Human S100A8/S100A9 Heterodimer 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.

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human S100A8/S100A9 Heterodimer Monoclonal Antibody (Catalog # MAB45703).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human S100A8/S100A9 Heterodimer DuoSet ELISA Kit (Catalog # DY8226-05) for convenient development of a sandwich ELISA or the Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit (Catalog # DS8900) for a complete optimized ELISA.

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon cancer tissue

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood monocytes (PBMC) fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer (Catalog # FC005)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *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.

Background: S100A8/S100A9 Heterodimer

S100A8 (also known as MRP8, Calgranulin A, and CP-10) and S100A9 (also known as MRP14 and Calgranulin B) are pro-inflammatory members of the S100 family of secreted calcium binding proteins (1, 2). They are up-regulated in neutrophils and monocytes at sites of inflammation (e.g. psoriasis, rheumatoid arthritis, cardiac ischemia) and are present at elevated concentrations in rheumatoid arthritis synovial fluid (3-5). The 10 kDa human S100A8 and 14 kDa S100A9 each contain two EF-hand calcium binding motifs. Human S100A8 shares 57% and 61% amino acid (aa) sequence identity with mouse and rat S100A8, respectively. Human S100A9 shares 57% and 62% amino acid sequence identity with mouse and rat S100A9, respectively (6, 7). In the presence of calcium or zinc, S100A8 and S100A9 associate into non-covalent homodimers and 34-35 kDa heterodimers with each other (8-10). The heterodimer additionally binds and sequesters manganese, thereby restricting the growth of Mn-dependent bacteria (11). The S100A8/A9 heterodimer exhibits functions beyond those performed by the individual proteins. These include binding to fatty acids such as arachidonic acid and promoting astrocyte proliferation (3, 12). S100A8, S100A9, and the heterodimer each promote neutrophil infiltration into sites of inflammation and inflammatory cytokine production by monocytes (4, 5, 9).

References

  1. Averill, M.M. et al. (2012) Arterioscler. Thromb. Vasc. Biol. 32:223.
  2. Vogl, T. et al. (2012) Int. J. Mol. Sci. 13:2893.
  3. Siegenthaler, G. et al. (1997) J. Biol. Chem. 272:9371.
  4. Sunahori, K. et al. (2006) Arthritis Res. Ther. 8:R69.
  5. Volz, H.C. et al. (2012) Basic Res. Cardiol. 107:250.
  6. Odink, K. et al. (1987) Nature 330:80.
  7. Dorin, J.R. et al. (1987) Nature 326:614.
  8. Teigelkamp, S. et al. (1991) J. Biol. Chem. 266:13462.
  9. Ryckman, C. et al. (2003) J. Immunol. 170:3233.
  10. Vogl, T. et al. (2006) Biochim. Biophys. Acta 1763:1298.
  11. Damo, S.M. et al. (2013) Proc. Natl. Acad. Sci. USA 110:3841.
  12. Ryu, M-J. et al. (2012) J. Biol. Chem. 287:22948.

Long Name

S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9

Alternate Names

Calprotectin

Entrez Gene IDs

6279 (Human)

Gene Symbol

S100A8

UniProt

P05109 (S100A8) and P06702 (S100A9)

Additional S100A8/S100A9 Heterodimer Products

Product Documents for Human S100A8/S100A9 Heterodimer Antibody

Product Datasheets

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human S100A8/S100A9 Heterodimer Antibody

For research use only

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