S100A8/S100A9 Heterodimer Antibody (1099F) [Unconjugated]

S100A8/S100A9 Heterodimer was detected in immersion fixed HL60 human promyelocytic leukemia cell line using Rabbit Anti-Human S100A8/S100A9 Heterodimer Monoclonal Antibody (Catalog # MAB45701) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm (punctate). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

S100A8/S100A9 Heterodimer was detected in immersion fixed paraffin-embedded sections of human lung cancer tissue using Rabbit Anti-Human S100A8/S100A9 Heterodimer Monoclonal Antibody (Catalog # MAB45701) at 10 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in tumor cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Syndecan-1/CD138 mRNA was detected (top image)in formalin-fixed paraffin-embedded tissue sections of human metastatic tonsilcancer tissue probed with ACD RNAScope® Probe (Catalog # 425271) and stainedusing ACD RNAscope® 2.5 HD Duplex Detection Reagents (red, Catalog # 322500).Adjacent tissue section (bottom image) was processed for immunohistochemistry using R&DSystems Rabbit Anti-Human S100A8/S100A9 Monoclonal Antibody (Catalog# MAB45701)at 5 ug/mL for 1 hour at room temperature followed by incubation with the Anti-RabbitIgG VisUCyte HRP Polymer Antibody (R&D Systems, Catalog # VC003) and DABchromogen (brown). Tissues were counterstained with hematoxylin (blue).

• Reactivity: Hu
• Applications: IHC, Dual ISH-IHC, ICC/IF
• Clone: 1099F
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

S100A8/S100A9 Heterodimer Antibody (1099F) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal 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 in direct ELISAs. In ELISAs, it detects recombinant human S100A8/S100A9 heterodimer but it does not detect recombinant human S100A8 or S100A9 monomers.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Dual RNAscope ISH-IHC 5-25 ug/mL
  • Immunocytochemistry 3-25 ug/mL
  • Immunohistochemistry 10-25 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: Sodium Azide
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

* Contains <0.1% Sodium Azide, which is not hazardous at this concentration according to GHS classifications. Refer to SDS for additional information and handling instructions.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for S100A8/S100A9 Heterodimer Antibody (1099F) [Unconjugated]

• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for S100A8/S100A9 Heterodimer Antibody (MAB45701)(2)

Publications using MAB45701

• Stempels FC, Jiang M, Warner HM et al. Giant worm-shaped ESCRT scaffolds surround actin-independent integrin clusters The Journal of cell biology 7/3/2023 [PMID: 37200023]
• F Real, A Zhu, B Huang, A Belmellat, A Sennepin, T Vogl, C Ransy, M Revol, R Arrigucci, A Lombès, J Roth, ML Gennaro, F Bouillaud, S Cristofari, M Bomsel S100A8-mediated metabolic adaptation controls HIV-1 persistence in macrophages in vivo Nature Communications, 2022-10-11;13(1):5956. 2022-10-11 [PMID: 36220814] (ICC, Human)

Bioinformatics

• Uniprot:
  • P05109 (S100A8) and P06702 (S100A9)()