S100A9 Antibody [Unconjugated]

Western blot shows lysate of mouse lung tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Mouse S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2065) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for S100A9 at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

S100A9 was detected in immersion fixed XB2 mouse teratoma keratinocyte cell line using 10 µg/mL Goat Anti-Mouse S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2065) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counte­rstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

S100A9 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2065) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

S100A9 was detected in perfusion fixed frozen sections of mouse spleen using Goat Anti-Mouse S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2065) at 0.3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in splenocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western lane view shows lysates of mouse lung tissue and mouse spleen tissue, loaded at 0.2 mg/mL. A specific band was detected for S100A9 at approximately 20 kDa (as indicated) using 2 µg/mL of Goat Anti-Mouse S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2065) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Mu
• Applications: WB, Simple Western, IHC, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

S100A9 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant mouse S100A9
  Ala2-Lys113
  Accession # P31725
• Specificity: Detects mouse S100A9 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 2% cross‑reactivity with recombinant mouse S100A10 and recombinant human S100B is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: S100A9
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 0.3-15 ug/mL
  • Simple Western 2 ug/mL
  • Western Blot 0.2 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: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

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

Alternate Names for S100A9 Antibody [Unconjugated]

• CAGBMigration inhibitory factor-related protein 14
• Calgranulin B
• calgranulin-B
• Calprotectin L1H subunit
• CFAGMRP-14,60B8AG
• CGLB
• L1AG
• LIAG
• MAC387
• MIF
• MRP-14
• MRP14Leukocyte L1 complex heavy chain
• NIF
• P14
• protein S100-A9
• S100 calcium binding protein A9 (calgranulin B)
• S100 calcium binding protein A9
• S100 calcium-binding protein A9 (calgranulin B)
• S100 calcium-binding protein A9
• S100A9

Background

S100A9, also known as calgranulin B, migration inhibitory factor-related protein 14 (MRP14), belongs to the S100 family of calcium binding proteins of the EF-hand type. It is highly expressed in myeloid cells and other cell types including keratinocytes, epithelial cells and endothelial cells. S100A9 is a cytosolic protein that can also be released into the extracellular milieu. It is associated with various inflammatory diseases.

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 S100A9 Antibody (AF2065)(21)

Publications using AF2065

• N Maehara, K Taniguchi, A Okuno, H Ando, A Hirota, Z Li, CT Wang, S Arai, T Miyazaki AIM/CD5L attenuates DAMPs in the injured brain and thereby ameliorates ischemic stroke Cell Reports, 2021;36(11):109693. 2021 [PMID: 34525359] (IHC, Mouse)
• MJ Kim, JY Kim, M Kang, MH Won, SH Hong, Y Her Reduced Fecal Calprotectin and Inflammation in a Murine Model of Atopic Dermatitis Following Probiotic Treatment Int J Mol Sci, 2020;21(11):. 2020 [PMID: 32486523] (Western Blot, Mouse)
• A Dufour, CL Bellac, U Eckhard, N Solis, T Klein, R Kappelhoff, N Fortelny, P Jobin, J Rozmus, J Mark, P Pavlidis, V Dive, SJ Barbour, CM Overall C-terminal truncation of IFN-? inhibits proinflammatory macrophage responses and is deficient in autoimmune disease Nat Commun, 2018;9(1):2416. 2018 [PMID: 29925830] (Western Blot, Human)
• E Källberg, S Tahvili, F Ivars, T Leanderson Induction of S100A9 homodimer formation in�vivo Biochem. Biophys. Res. Commun., 2018;0(0):. 2018 [PMID: 29679568] (Western Blot, Mouse)
• Beneficial Effects of Sodium Phenylbutyrate Administration During Infection with Salmonella enterica serovar Typhimurium Infect Immun, 2016;0(0):. 2016 [PMID: 27382022] (Western Blot, Mouse)
• Influenza Virus Affects Intestinal Microbiota and Secondary Salmonella Infection in the Gut through Type I Interferons PLoS Pathog, 2016;12(5):e1005572. 2016 [PMID: 27149619] (Western Blot, Mouse)
• Oncostatin M overexpression induces skin inflammation but is not required in the mouse model of imiquimod-induced psoriasis-like inflammation Eur J Immunol, 2016;0(0):. 2016 [PMID: 27122058] (IHC-Fr, Mouse)
• Armstrong P, Kirkby N, Chan M, Finsterbusch M, Hogg N, Nourshargh S, Warner T Novel whole blood assay for phenotyping platelet reactivity in mice identifies ICAM-1 as a mediator of platelet-monocyte interaction. Blood, 2015;126(10):e11-8. 2015 [PMID: 26215112] (Confocal Microscopy, Mouse)
• Thomsen M, Bakiri L, Hasenfuss S, Wu H, Morente M, Wagner E Loss of JUNB/AP-1 promotes invasive prostate cancer. Cell Death Differ, 2015;22(4):574-82. 2015 [PMID: 25526087] (Western Blot, Mouse)
• Jiao , Jingjing, Dragomir , Ana-Cris, Kocabayoglu , Peri, Rahman , Adeeb H, Chow , Andrew, Hashimoto , Daigo, Leboeuf , Marylene, Kraus , Thomas, Moran , Thomas, Carrasco-Avino , Gonzalo, Friedman , Scott L, Merad , Miriam, Aloman , Costica Central role of conventional dendritic cells in regulation of bone marrow release and survival of neutrophils. J Immunol, 2014;192(7):3374-82. 2014 [PMID: 24591364] (IHC, Mouse)
• Kim H, Chang K, Ha T, Kim J, Ha S, Shin K, Moon C, Nacken W, Kim H, Suh Y S100A9 knockout decreases the memory impairment and neuropathology in crossbreed mice of Tg2576 and S100A9 knockout mice model. PLoS ONE, 2014;9(2):e88924. 2014 [PMID: 24586443] (IHC, Mouse)
• Behnsen J, Jellbauer S, Wong C, Edwards R, George M, Ouyang W, Raffatellu M The cytokine IL-22 promotes pathogen colonization by suppressing related commensal bacteria. Immunity, 2014;40(2):262-73. 2014 [PMID: 24508234] (Western Blot, Mouse)
• Yano J, Palmer G, Eberle K, Peters B, Vogl T, McKenzie A, Fidel P Vaginal epithelial cell-derived S100 alarmins induced by Candida albicans via pattern recognition receptor interactions are sufficient but not necessary for the acute neutrophil response during experimental vaginal candidiasis. , 2014;0(0):. 2014 [PMID: 24478092] (Neutralization, Mouse)
• Sade-Feldman M, Kanterman J, Ish-Shalom E, Elnekave M, Horwitz E, Baniyash M Tumor necrosis factor-alpha blocks differentiation and enhances suppressive activity of immature myeloid cells during chronic inflammation. Immunity, 2013;38(3):541-54. 2013 [PMID: 23477736] (Western Blot, Mouse)
• Yano , Junko, Kolls , Jay K, Happel , Kyle I, Wormley , Floyd, Wozniak , Karen L, Fidel , Paul L J The Acute Neutrophil Response Mediated by S100 Alarmins during Vaginal Candida Infections Is Independent of the Th17-Pathway. PLoS ONE, 2012;7(9):e46311. 2012 [PMID: 23050010] (ELISA Development, Mouse)
• Gonzalez-Lopez A, Aguirre A, Lopez-Alonso I, Amado L, Astudillo A, Fernandez-Garcia MS, Suarez MF, Batalla-Solis E, Colado E, Albaiceta GM MMP-8 Deficiency Increases TLR/RAGE Ligands S100A8 and S100A9 and Exacerbates Lung Inflammation during Endotoxemia. PLoS ONE, 2012;7(6):e39940. 2012 [PMID: 22768176] (Western Blot, Mouse)
• Fijneman RJ, de Wit M, Pourghiasian M, Piersma SR, Pham TV, Warmoes MO, Lavaei M, Piso C, Smit F, Delis-van Diemen PM, van Turenhout ST, Terhaar sive Droste JS, Mulder CJ, Blankenstein MA, Robanus-Maandag EC, Smits R, Fodde R, van Hinsbergh VW, Meijer GA, Jimenez CR Proximal fluid proteome profiling of mouse colon tumors reveals biomarkers for early diagnosis of human colorectal cancer. Clin. Cancer Res., 2012;18(9):2613-24. 2012 [PMID: 22351690] (IHC-P, Mouse)
• Taccioli , C, Chen , H, Jiang , Y, Liu , X P, Huang , K, Smalley , K J, Farber , J L, Croce , C M, Fong , L Y Dietary zinc deficiency fuels esophageal cancer development by inducing a distinct inflammatory signature. Oncogene, 2012;31(42):4550-8. 2012 [PMID: 22179833] (Western Blot, Rat)
• Cuervo H, Guerrero NA, Carbajosa S, Beschin A, De Baetselier P, Girones N, Fresno M Myeloid-Derived Suppressor Cells Infiltrate the Heart in Acute Trypanosoma cruzi Infection. J. Immunol., 2011;187(5):2656-65. 2011 [PMID: 21804013] (Western Blot, Mouse)
• Farris SD, Hu JH, Krishnan R, Emery I, Chu T, DU L, Kremen M, Dichek HL, Gold E, Ramsey SA, Dichek DA Mechanisms of Urokinase Plasminogen Activator (uPA)-mediated Atherosclerosis: ROLE OF THE uPA RECEPTOR AND S100A8/A9 PROTEINS. J. Biol. Chem., 2011;286(25):22665-77. 2011 [PMID: 21536666] (Western Blot, Mouse)
• Mares CA, Ojeda SS, Morris EG, Li Q, Teale JM Initial delay in the immune response to Francisella tularensis is followed by hypercytokinemia characteristic of severe sepsis and correlating with upregulation and release of damage-associated molecular patterns. Infect. Immun., 2008;76(7):3001-10. 2008 [PMID: 18411294] (Functional Assay, Mouse)

Reviews for S100A9 Antibody (AF2065) (2)

Average Rating: 4

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Mouse.

reviewed by: Anonymous - IHC-Fr Mouse 01/06/2015

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: See PMID 21228116
• Species: Mouse

reviewed by: Anonymous - IF Mouse 01/06/2015

Summary

• Application: Immunofluorescence
• Sample Tested: See PMID 23481567
• Species: Mouse

FAQs for S100A9 Antibody (AF2065). (Showing 1 - 2 of 2 FAQs).

1. Have any of your S100A9 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 S100A9 antibodies for use in blocking or neutralizing assays. If you were planning on testing any of our S100A9 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 <a href="http://www.novusbio.com/support/innovators-reward.html" target="_self">Innovators Reward Program</a> 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: S100A9
• Uniprot:
  • P31725()