Human IL-15 Antibody (34559)

R&D Systems | Catalog # MAB2471

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Neutralization, Intracellular Staining by Flow Cytometry, CyTOF-ready

Cited:

Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Blocking, Cell Culture

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 34559
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Product Specifications

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, no cross-reactivity with recombinant human (rh) IL-2, recombinant mouse IL-15, or rhIL-21 is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Endotoxin Level

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

Scientific Data Images for Human IL-15 Antibody (34559)

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

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

Recombinant Human IL-15 (Catalog # 247-ILB) stimulates proliferation in the MO7e human megakaryocytic leukemic cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Human IL-15 (5 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-15 Monoclonal Antibody (Catalog # MAB2471). The ND50 is typically 0.1-0.5 µg/mL.

Detection of IL‑15 in Human peripheral blood mononuclear cells (PBMCs) treated with 1µg/mL LPS for 16 hrs by Flow Cytometry

Human peripheral blood mononuclear cells (PBMCs) treated with 1µg/mL LPS for 16 hrs were stained with Mouse Anti-Human IL‑15 Monoclonal Antibody (Catalog # MAB2471, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram) followed by Fluorescein-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0103B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Applications for Human IL-15 Antibody (34559)

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: See below

Western Blot

1 µg/mL
Sample: Recombinant Human IL‑15 (Catalog # 247-IL)

Neutralization

Measured by its ability to neutralize IL‑15-induced proliferation in the MO7e human megakaryocytic leukemic cell line. Avanzi, G. et al. (1988) Br. J. Haematol. 69:359. The Neutralization Dose (ND50) is typically 0.1-0.5 µg/mL in the presence of 5 ng/mL Recombinant Human IL‑15.

Reviewed Applications

Read 3 reviews rated 5 using MAB2471 in the following applications:

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 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).

References

  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.

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

Additional IL-15 Products

Product Documents for Human IL-15 Antibody (34559)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human IL-15 Antibody (34559)

For research use only

Citations for Human IL-15 Antibody (34559)

Customer Reviews for Human IL-15 Antibody (34559) (3)

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3 Customer Ratings
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Showing  1 - 3 of 3 reviews Showing All
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  • Human IL-15 Antibody
    Name: Anonymous
    Application: I use the GBM cells lines which overexpression with mIL15, and test by the MAB247 IL15 antibody, the immunostaining results shows strong signal of IL15 located on the cell membrane, I am excited about that.
    Sample Tested: A172 human glioblastoma cell line
    Species: Human
    Verified Customer | Posted 05/01/2024
    Human IL-15 Antibody (34559) MAB2471
  • Human IL-15 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: NK cells
    Species: Human
    Verified Customer | Posted 09/30/2021
    Human IL-15 Antibody (34559) MAB2471
  • Name: Anonymous
    Application: ELISA
    Sample Tested: Recombinant protein
    Species: Human
    Verified Customer | Posted 02/08/2021
    Human IL-15 Antibody (34559) MAB2471

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FAQs for Human IL-15 Antibody (34559)

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  • Q: What concentration of LPS was used to stimulate PBMCs prior to staining with Human IL-15 Antibody, Catalog #s MAB2471 and IC2471?

    A: PBMCs were stimulated overnight with 1 μg/mL LPS prior to staining with Human IL-15 Antibody. 3mM Monesin was added for the last 3-4 hours of stimulation.

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