Detects mouse IL-15 in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with recombinant human IL‑15 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse IL-15 Antibody

Cell Proliferation Induced by IL‑15 and Neutralization by Mouse IL‑15 Antibody.

Recombinant Mouse IL-15 (Catalog # 447-ML) stimulates proliferation in the CTLL-2 mouse cytotoxic T cell line in a dose-dependent manner (orange line) as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Mouse IL-15 (30 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse IL-15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF447). The ND₅₀ is typically 0.4-2.4 µg/mL.

IL-15 in Mouse Intestine.

IL-15 was detected in immersion fixed frozen sections of mouse intestine (Peyer patch) using Goat Anti-Mouse IL-15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF447) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm of lymphocytes in villi. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of IL-15 in Mouse Intestine.

Formalin-fixed paraffin-embedded tissue sections of mouse intestine were probed for IL-15 mRNA (ACD RNAScope Probe, catalog # 440651; Fast Red chromogen, ACD catalog # 322360). Adjacent tissue section was processed for immunohistochemistry using goat anti-mouse IL-15 polyclonal antibody (R&D Systems catalog # AF447) at 1ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

Detection of Mouse IL-15 by Western Blot

Eugenol promotes IL-15 level through TRPV1-mediated CaN/NFATc1 signaling pathway.C2C12 myotubes were treated by 0–200 eugenol for 1 day after 4 days of differentiation. (A) The effect of eugenol on Il15 mRNA expression in C2C12 myotubes. (B) The effect of eugenol on IL-15 protein expression in the C2C12 cell medium. Coomassie staining as loading control. (C, D) C2C12 myotubes were treated by 25 μM eugenol & 1 μM TRPV1 inhibitor AMG-517 or 0.5 μM calcineurin (CaN) inhibitor cyclosporine A (CsA) for 1 day after 4 days of differentiation. The mRNA & protein expression of IL-15. N=4 per group. One-way ANOVA test was used to determine statistical significance. *p<0.05, **p<0.01, & ***p<0.001.Figure 9—source data 1.IL-15 mRNA expression in C2C12 myotubes (Figure 9A).Figure 9—source data 2.Original files for the WB analysis (Figure 9B).Figure 9—source data 3.PDF containing Figure 9B & original scans of the relevant WB analysis, with cropped areas. Figure 9—source data 4.IL-15 mRNA expression in C2C12 myotubes (Figure 9C).Figure 9—source data 5.Original files for the WB analysis (Figure 9D).Figure 9—source data 6.PDF containing Figure 9D & original scans of the relevant WB analysis, with cropped areas.IL-15 mRNA expression in C2C12 myotubes (Figure 9A).Original files for the WB analysis (Figure 9B).PDF containing Figure 9B & original scans of the relevant WB analysis, with cropped areas.IL-15 mRNA expression in C2C12 myotubes (Figure 9C).Original files for the WB analysis (Figure 9D).PDF containing Figure 9D & original scans of the relevant WB analysis, with cropped areas. Representative immunofluorescence images of IL-15.IL-15 (green fluorescence) & 4,6-diamidino-2-phenylindole (DAPI) (blue fluorescence). Magnification: ×200. Image collected & cropped by CiteAb from the following open publication (https://elifesciences.org/articles/90724), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse IL-15 by Western Blot

Eugenol promotes the expression and secretion of IL-15 in skeletal muscle of mice. (E) The protein expression of slow myosin heavy chain (MyHC), fast MyHC, and IL-15. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/90724), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse IL-15 by Western Blot

The myokine IL-15 expression depends on CaN/NFATc1 signaling pathway. (B–D) C2C12 myotubes were treated by 0.5 μM A23187 and 0.5 μM cyclosporine A (CsA) for 16 hr after 2 days of differentiation. The protein expression of CnA, NFATc1, and IL-15. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/90724), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse IL-15 by Western Blot

Mouse IL-15 ELISA Standard Curve

Recombinant Mouse IL-15 (Catalog # 447-ML) was serially diluted and captured by Rat Anti-Mouse IL-15 Monoclonal Antibody (Catalog # MAB447) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse IL-15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF447) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

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Applications for Mouse IL-15 Antibody

Application

Recommended Usage

Dual RNAscope ISH-IHC Compatible

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of mouse intestine

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse intestine (Peyer patch)

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse IL-15 (Catalog # 447-ML)

Neutralization

Measured by its ability to neutralize IL‑15-induced proliferation in the CTLL‑2 mouse cytotoxic T cell line. Avanzi, G. et al. (1988) Br. J. Haematol. 69:359. The Neutralization Dose (ND₅₀) is typically 0.4-2.4 µg/mL in the presence of 30 ng/mL Recombinant Mouse IL‑15.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in 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: IL-15

Interleukin 15 (IL-15) is a widely expressed 14 kDa cytokine that is structurally and functionally related to IL-2 (1-3). Mature mouse IL-15 shares 70% and 96% amino acid sequence identity with human and rat IL-15, respectively. 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 ( gammac) 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/ gammac 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).

References

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

Long Name

Interleukin 15

Alternate Names

IL15

Entrez Gene IDs

3600 (Human); 16168 (Mouse); 25670 (Rat); 102119613 (Cynomolgus Monkey); 493682 (Feline)

Gene Symbol

IL15

UniProt

P48346

Additional IL-15 Products

Product Documents for Mouse IL-15 Antibody

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COA

SDS

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Product Specific Notices for Mouse IL-15 Antibody

For research use only

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