S100A8 Antibody (749916)

Western blot shows lysates of HL-60 human acute promyelocytic leukemia cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human S100A8 Monoclonal Antibody (Catalog # MAB4570) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for S100A8 at approximately 11 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

S100A8 was detected in immersion fixed paraffin-embedded sections of human lung cancer tissue using Mouse Anti-Human S100A8 Monoclonal Antibody (Catalog # MAB4570) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei and cytoplasm of cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Quantitative PCR and immunoblotting of five candidate gene products in DLBCL and LCL lines. (A) Reverse transcriptase and real-time PCR analysis of mRNAs for BMP8A, CCR6, HOXA9, NANOG and S100A8 in LCL and DLBCL (HBL-2, HT, SU-DHL-5, U2932 and U2940) cell lines. Data were normalized by the amount of GAPDH mRNA, expressed relative to the corresponding value for LCL, and are shown as means ± SD from triplicate data. (B) After quantification of Western blot intensity, the relative expression levels of each protein are shown with normalization of GAPDH. U2940 cells have the highest expression of the stem cell genes, HOXA9 and NANOG. Molecular weight listed in parenthesis. Error bars represent the standard error of the mean of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, Student t-test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33288848), licensed under a CC-BY license. Not internally tested by R&D Systems.

Quantitative PCR and immunoblotting of five candidate gene products in DLBCL and LCL lines. (A) Reverse transcriptase and real-time PCR analysis of mRNAs for BMP8A, CCR6, HOXA9, NANOG and S100A8 in LCL and DLBCL (HBL-2, HT, SU-DHL-5, U2932 and U2940) cell lines. Data were normalized by the amount of GAPDH mRNA, expressed relative to the corresponding value for LCL, and are shown as means ± SD from triplicate data. (B) After quantification of Western blot intensity, the relative expression levels of each protein are shown with normalization of GAPDH. U2940 cells have the highest expression of the stem cell genes, HOXA9 and NANOG. Molecular weight listed in parenthesis. Error bars represent the standard error of the mean of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, Student t-test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33288848), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IHC
• Clone: 749916
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

S100A8 Antibody (749916) Summary

• Immunogen: E. coli-derived recombinant human S100A8
  Met1-Glu93
  Accession # P05109
• Specificity: Detects human S100A8 in direct ELISAs. In direct ELISAs, 100% cross-reactivity with recombinant human (rh) S100A8/A9 is observed and no cross-reactivity with rhS100A9, recombinant mouse (rm) S100A8, rmS100A9, or rmS100A8/A9 is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: S100A8
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 8-25 ug/mL
  • Western Blot 2 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: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

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

Alternate Names for S100A8 Antibody (749916)

• 60B8AG
• CAGACP-10
• Calgranulin A
• calgranulin-A
• Calprotectin L1L subunit
• CFAG
• CFAGL1Ag
• CGLA
• Cystic fibrosis antigen
• Leukocyte L1 complex light chain
• MA387
• MIF
• Migration inhibitory factor-related protein 8
• MRP8
• MRP-8
• MRP8S100 calcium binding protein A8 (calgranulin A)
• NIF
• P8
• protein S100-A8
• S100 calcium binding protein A8
• S100 calcium-binding protein A8 (calgranulin A)
• S100 calcium-binding protein A8
• S100A8
• Urinary stone protein band A

Background

S100A8 (also known as MRP8 and calgranulin A) is a 10 kDa member of the S100 family, EF-hand superfamily of Ca-binding proteins. It is produced by neutrophils and monocytes, and forms Ca‑dependent heterodimer/heterotetramer complexes (termed calprotectin) with S100A9. It functions both intracellularly and extracellularly, where it binds to RAGE and CD36. Human S100A8 is 93 amino acids (aa) in length. It contains two EF-hand motifs (aa 12-47 and 46-81) and one high-affinity Ca‑binding site (aa 59-70). There may be one splice form that shows a 15 aa substitution for the C-terminal 14 amino acids. Although mouse S100A8 is cleaved by MMP‑2 after Asn21, it is unclear if human S100A8 is susceptible. Full-length human S100A8 is 57% and 74% aa identical to mouse and canine S100A8, respectively.

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 Antibody (MAB4570)(11)

Publications using MAB4570

• Mathieu Le Boulvais Børkja, Miriam S. Giambelluca, Borgny Ytterhus, Wenche S. Prestvik, Geir Bjørkøy, Anna M Bofin S100A8 gene copy number and protein expression in breast cancer: associations with proliferation, histopathological grade and molecular subtypes Breast Cancer Research and Treatment 9/1/2023 [PMID: 37450087]
• K Asleh, GL Negri, SE Spencer Mi, S Colborne, CS Hughes, XQ Wang, D Gao, CB Gilks, SKL Chia, TO Nielsen, GB Morin Proteomic analysis of archival breast cancer clinical specimens identifies biological subtypes with distinct survival outcomes Nature Communications, 2022-02-16;13(1):896. 2022-02-16 [PMID: 35173148] (IHC, Human)
• EGG Sprenkeler, J Zandstra, ND van Kleef, I Goetschalc, B Verstegen, CEM Aarts, H Janssen, ATJ Tool, G van Mierlo, R van Brugge, I Jongerius, TW Kuijpers S100A8/A9 Is a Marker for the Release of Neutrophil Extracellular Traps and Induces Neutrophil Activation Cells, 2022-01-11;11(2):. 2022-01-11 [PMID: 35053354] (ICC, Human)
• Ingunn Bakke, Gunnar Andreas Walaas, Torunn Bruland, Elin Synnøve Røyset, Atle van Beelen Granlund, Celia Escudero-Hernández et al. Mucosal and faecal neutrophil gelatinase-associated lipocalin as potential biomarkers for collagenous colitis Journal of Gastroenterology 10/1/2021 [PMID: 34414506]
• Ingunn Bakke, Gunnar Andreas Walaas, Torunn Bruland, Elin Synnøve Røyset, Atle van Beelen Granlund, Celia Escudero-Hernández et al. Mucosal and faecal neutrophil gelatinase-associated lipocalin as potential biomarkers for collagenous colitis Journal of Gastroenterology 2021-10-01 [PMID: 34414506] (Immunohistochemistry, Human)
• KC Chang, RY Chen, YC Wang, LY Hung, LJ Medeiros, YP Chen, TY Chen, JC Yang, PM Chiang Stem cell characteristics promote aggressiveness of diffuse large B-cell lymphoma Scientific Reports, 2020-12-07;10(1):21342. 2020-12-07 [PMID: 33288848] (Western Blot, Human)
• J Schmidt, B Kajtár, K Juhász, M Péter, T Járai, A Burián, L Kereskai, I Gerlinger, T Tornóczki, G Balogh, L Vígh, L Márk, Z Balogi Lipid and protein tumor markers for head and neck squamous cell carcinoma identified by imaging mass spectrometry Oncotarget, 2020-07-14;11(28):2702-2717. 2020-07-14 [PMID: 32733643] (IHC, Human)
• Dujuan Wang, Guohong Liu, Balu Wu, Li Chen, Lihua Zeng, Yunbao Pan Clinical Significance of Elevated S100A8 Expression in Breast Cancer Patients Frontiers in Oncology 11/5/2018 [PMID: 30456203]
• Janet P.C. Wong, Ran Wei, Peng Lyu, Olivia L.H. Tong, Shu Dong Zhang, Qing Wen et al. Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer International Journal of Biological Sciences 11/1/2017 [PMID: 29209142]
• Janet P.C. Wong, Ran Wei, Peng Lyu, Olivia L.H. Tong, Shu Dong Zhang, Qing Wen et al. Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer International Journal of Biological Sciences 2017-11-01 [PMID: 29209142] (Western Blot, Human)
• Sandra Founds, Xuemei Zeng, David Lykins, James M. Roberts Developing Potential Candidates of Preclinical Preeclampsia International Journal of Molecular Sciences 11/13/2015 [PMID: 26580600]

FAQs for S100A8 Antibody (MAB4570). (Showing 1 - 2 of 2 FAQs).

1. Have any of your MRP8 antibodies been tested for use in neutralization, or blocking assays, on human samples?
   • Unfortunately at this time we have not tested, or received customer feedback on our MMP8 antibodies for use in blocking or neutralizing assays. If you were planning on testing any of our MMP8 antibodies for neutralizing or blocking capabilities, we would recommend our Innovator’s Reward Program. Under the terms of this program we would be happy to provide a credit for a free vial in exchange for new data on this previously untested or reported application. Please submit this data in the form of an online review. Additional Innovators Reward Program information can be found using this link.
2. We would like to stain paraffin embedded human intestinal tissue for Calprotectin. One question will be which cells express the majority of Calprotectin - invading neutrophils vs epithelium for example. The antibody list you provide is quite extensive. Which antibody would you recommend for our purpose? Which tissue would you recommend as positive control.
   • Nine of our antibodies to Calprotectin have been validated for IHC-P with human tissue, and you can see all of these products. A number of our IHC images were generated using human spleen samples, and as such this may be a good choice of a positive control tissue. We do sell a human spleen slide product, which is suitable for IHC-P. Unfortunately I do not have an in-depth knowledge of Calprotectin, however the image shown for our antibody with catalogue number NBP1-02826 demonstrates clear staining of neutrophils. The following paper also suggests that the protein is abundant in neutrophils: PMID 11435495.

Bioinformatics

• Gene Symbol: S100A8
• Uniprot:
  • P05109()