IL-15 Antibody [Unconjugated]

IL‑15 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human IL‑15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF315) at 10 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-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Recombinant Human IL‑15 (Catalog # 247-IL) stimulates proliferation in the MO7e human megakaryocytic leukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL‑15 (10 ) is neutralized (green line) by increasing concen­trations of Goat Anti-Human IL‑15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF315). The ND50 is typically 0.5-1.5 µg/mL.

• Reactivity: Hu
• Applications: WB, IHC, B/N
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

IL-15 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human IL-15
  Asn49-Ser162
  Accession # P40933
• Specificity: Detects human IL-15 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant mouse IL‑15 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: IL15
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Neutralization 0.5-1.5 ug/mL
  • Western Blot 0.1 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 IL-15 Antibody [Unconjugated]

• IL15
• IL-15
• IL-15MGC9721
• interleukin 15
• interleukin-15

Background

Interleukin 15 (IL-15) is a widely expressed 14 kDa cytokine that is structurally and functionally related to IL-2 (1-3). Mature human IL-15 shares 70% amino acid sequence identity with mouse and rat IL-15. Alternate splicing generates isoforms of IL-15 with either a long or short signal peptide (LSP or SSP), and the SSP isoform is retained intracellularly (4). IL-15 binds with high affinity to IL-15 R alpha (5). It binds with lower affinity to a complex of IL-2 R beta and the common gamma chain ( gamma c) which are also subunits of the IL-2 receptor complex (1, 6). IL-15 associates with IL-15 R alpha in the endoplasmic reticulum, and this complex is expressed on the cell surface (7, 8). The dominant mechanism of IL-15 action is known as transpresentation in which IL-15 and IL-15 R alpha are coordinately expressed on the surface of one cell and interact with complexes of IL-2 R beta / gamma c on adjacent cells (9). This enables cells to respond to IL-15 even if they do not express IL-15 R alpha (8, 10). Soluble IL-15-binding forms of IL-15 R alpha can be generated by proteolytic shedding or alternate splicing (11-13). These molecules retain the ability to bind tightly to IL-15 and can either inhibit or augment IL-15 function (5, 12, 13). Consistent with its shared use of IL-2 receptor subunits, IL-15 induces IL-2-like effects in lymphocyte development and homeostasis (3). It is particularly important for the maintenance and activation of NK cells and CD8⁺ memory T cells (3). IL-15 also exerts pleiotropic effects on other hematopoietic cells and non-immune cells (2). Ligation of membrane-associated IL-15/IL-15 R alpha complexes induces reverse signaling that promotes cellular adhesion, tyrosine phosphorylation of intracellular proteins, and cytokine secretion by the IL-15/IL-15 R alpha expressing cells (14, 15).

1. Grabstein, K. et al. (1994) Science 264:965.
2. Budagian, V. et al. (2006) Cytokine Growth Factor Rev. 17:259.
3. Ma, A. et al. (2006) Annu. Rev. Immunol. 24:657.
4. Tagaya, Y. et al. (1997) Proc. Natl. Acad. Sci. USA 94:14444.
5. Giri, J.G. et al. (1995) EMBO 14:3654.
6. Giri, J. et al. (1994) EMBO J. 13:2822.
7. Duitman, E.H. et al. (2008) Mol. Cell. Biol. 28:4851.
8. Dubois, S. et al. (2002) Immunity 17:537.
9. Stonier, S.W. and K.S. Schluns (2010) Immunol. Lett. 127:85.
10. Burkett, P.R. et al. (2004) J. Exp. Med. 200:825.
11. Budagian, V. et al. (2004) J. Biol. Chem. 279:40368.
12. Mortier, E. et al. (2004) J. Immunol. 173:1681.
13. Bulanova, E. et al. (2007) J. Biol. Chem. 282:13167.
14. Budagian, V. et al. (2004) J. Biol. Chem. 279:42192.
15. Neely, G.G. et al. (2004) J. Immunol. 172:4225.

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 IL-15 Antibody (AF315)(11)

Publications using AF315

• JS Miller, C Morishima, DG McNeel, MR Patel, HE Kohrt, JA Thompson, PM Sondel, HA Wakelee, ML Disis, JC Kaiser, MA Cheever, H Streicher, SP Creekmore, TA Waldmann, KC Conlon A First-in-Human Phase 1 Study of Subcutaneous Outpatient Recombinant Human IL-15 (rhIL-15) in Adults with Advanced Solid Tumors Clin. Cancer Res., 2017-12-04;0(0):. 2017-12-04 [PMID: 29203590]
• AP Vicari, AM Schoepfer, B Meresse, L Goffin, O Léger, S Josserand, N Guégan, S Yousefi, A Straumann, N Cerf-Bensu, HU Simon, Y Chvatchko Discovery and characterization of a novel humanized anti-IL-15 antibody and its relevance for the treatment of refractory celiac disease and eosinophilic esophagitis MAbs, 2017-06-05;0(0):0. 2017-06-05 [PMID: 28581883]
• Dionysios C. Watson, Defne Bayik, Avinash Srivatsan, Cristina Bergamaschi, Antonio Valentin, Gang Niu et al. Efficient production and enhanced tumor delivery of engineered extracellular vesicles Biomaterials 2016-10-01 [PMID: 27522254] (Western Blot, Human)
• M. Thaysen-Andersen, E. Chertova, C. Bergamaschi, E. S. X. Moh, O. Chertov, J. Roser et al. Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation Glycoconjugate Journal 2016-06-01 [PMID: 26563299]
• Mongini P, Gupta R, Boyle E, Nieto J, Lee H, Stein J, Bandovic J, Stankovic T, Barrientos J, Kolitz J, Allen S, Rai K, Chu C, Chiorazzi N TLR-9 and IL-15 Synergy Promotes the In Vitro Clonal Expansion of Chronic Lymphocytic Leukemia B Cells. J Immunol, 2015-07-01;195(3):901-23. 2015-07-01 [PMID: 26136429] (IHC-P, Human)
• Broux B, Mizee M, Vanheusden M, van der Pol S, van Horssen J, Van Wijmeersch B, Somers V, de Vries H, Stinissen P, Hellings N IL-15 amplifies the pathogenic properties of CD4+CD28- T cells in multiple sclerosis. J Immunol, 2015-01-23;194(5):2099-109. 2015-01-23 [PMID: 25617471] (IHC-Fr, Human)
• Bergamaschi C, Kulkarni V, Rosati M, Alicea C, Jalah R, Chen S, Bear J, Sardesai N, Valentin A, Felber B, Pavlakis G Intramuscular delivery of heterodimeric IL-15 DNA in macaques produces systemic levels of bioactive cytokine inducing proliferation of NK and T cells. Gene Ther, 2014-10-02;22(1):76-86. 2014-10-02 [PMID: 25273353] (Western Blot, Human)
• Marcondes MC, Burdo TH, Sopper S, Huitron-Resendiz S, Lanigan C, Watry D, Flynn C, Zandonatti M, Fox HS Enrichment and persistence of virus-specific CTL in the brain of simian immunodeficiency virus-infected monkeys is associated with a unique cytokine environment. J. Immunol., 2007-05-01;178(9):5812-9. 2007-05-01 [PMID: 17442965] (IHC-P, Primate - Macaca mulatta (Rhesus Macaque))
• Vasir B, Avigan D, Wu Z, Crawford K, Turnquist S, Ren J, Kufe D Dendritic cells induce MUC1 expression and polarization on human T cells by an IL-7-dependent mechanism. J. Immunol., 2005-02-15;174(4):2376-86. 2005-02-15 [PMID: 15699174] (Neutralization, Human)
• Dean GA, Barger A, Lavoy A Cloning and expression of feline interleukin 15. Cytokine, 2005-01-21;29(2):77-83. 2005-01-21 [PMID: 15598442] (ICC, Western Blot, Feline)
• Vankayalapati R, Klucar P, Wizel B, Weis SE, Samten B, Safi H, Shams H, Barnes PF NK cells regulate CD8+ T cell effector function in response to an intracellular pathogen. J. Immunol., 2004-01-01;172(1):130-7. 2004-01-01 [PMID: 14688318] (Neutralization, Human)

Bioinformatics

• Gene Symbol: IL15
• Uniprot:
  • P40933()