Human S100A8/S100A9 Heterodimer Antibody

Catalog #: MAB45702 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB45702-100
MAB45702-SP

S100A8/S100A9 Heterodimer in Human Colon Cancer Tissue.

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Human S100A8/S100A9 Heterodimer Antibody Summary

Species Reactivity

Human

Specificity

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

Source

Monoclonal Mouse IgG₁ Clone # 900027

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

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

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

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

See below

CyTOF-ready

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

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunohistochemistry

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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.

Intracellular Staining by Flow Cytometry

Detection of S100A8/A9 antibody in Human PBMCs antibody by Flow Cytometry.

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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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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

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

Long Name

S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9

Entrez Gene IDs

6279 (Human)

Alternate Names

60B8AG; CAGA; calgranulin A; Calprotectin L1L subunit; Calprotectin; CFAG; CGLA; CP-10; Cystic fibrosis antigen; L1AG; Leukocyte L1 complex light chain; MA387; MIF; Migration inhibitory factor-related protein 8; MRP8; NIF; p8; protein S100-A8; S100-A8; S100A8/S100A9 Heterodimer; Urinary stone protein band A