IL-15 Antibody (34505)

R&D Systems | Catalog # MAB647

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human, Primate

Cited:

Human, Mouse

Applications

Validated:

ELISA Capture (Matched Antibody Pair), Neutralization

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Bioassay, ELISA Capture, ELISA Development, Luminex Development

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 34505
View all IL-15 Primary Antibodies »

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 ELISAs, no cross-reactivity with recombinant human (rh) IL-1 alpha, -1 beta, -2, -3, -4, -5, -6, -7, -8, -9, rmIL-1 alpha, -1 beta, -3, -4, -5, -6, -7, -9, or -13 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 IL-15 Antibody (34505)

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) induces proliferation in the MO7e human megakaryocytic leukemic cell line in a dose-dependent manner (orange line), as measured by the Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Human IL-15 (3 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human/Primate IL-15 Monoclonal Antibody (Catalog # MAB647). The ND50 is typically 4-20 ng/mL.
Detection of Human IL-15 by Flow Cytometry

Detection of Human IL-15 by Flow Cytometry

Reciprocal activation between NK&THP-1 cells, the role of IL-15&IFN-gamma. (c) IFN-gamma increases IL-15 membrane expression in THP-1 cells. Cells incubated with IFN-gamma (100 ng/ml), TNF (100 ng/ml) or IL-1 (100 ng/ml) for 24 hours&then the membrane-bound IL-15 (mIL-15) was measured by indirect immunofluorescence&flow cytometry. A representative experiment of the five performed is shown. Grey histograms represent mIL-15 on stimulated THP-1 cells, solid-line histograms represent basal mIL-15 expression,&the dotted-line histogram is the negative control. Image collected & cropped by CiteAb from the following open publication (https://arthritis-research.biomedcentral.com/articles/10.1186/ar1955), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-15 by Flow Cytometry

Detection of Human IL-15 by Flow Cytometry

Reciprocal activation between NK&THP-1 cells, the role of IL-15&IFN-gamma. (a) IL-15& beta 2 integrins are involved in the intercellular contact with THP-1 cells that induces the expression of CD69 in natural killer (NK) cells. Peripheral blood lymphocytes (PBL) co-cultured with THP-1 cells (10:1 ratio of PBL to THP-1) for 24 hours in medium alone or in the presence of an anti-beta 2 integrin mAb (Lia3/2) or an anti-IL-15 mAb (MAB647). As a control, both cell lines separated by a 0.4 μm pore transwell. A representative experiment of the five performed is shown. The histograms represent the CD69 expression in CD56+ cells in the medium (grey histogram in all panels) or under the different conditions (solid black line in each panel); a negative control is also shown (dotted histogram in all panels). Image collected & cropped by CiteAb from the following open publication (https://arthritis-research.biomedcentral.com/articles/10.1186/ar1955), licensed under a CC-BY license. Not internally tested by R&D Systems.
Human IL-15 ELISA Standard Curve

Human IL-15 ELISA Standard Curve

Human IL-15 was serially diluted and captured by Mouse Anti-Human/Primate IL‑15 Monoclonal Antibody (Catalog # MAB647) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human IL‑15 Monoclonal Antibody (Catalog # MAB247) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for IL-15 Antibody (34505)

Application
Recommended Usage

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 4-20 ng/mL in the presence of 3 ng/mL Recombinant Human IL‑15.

Human/Primate IL-15 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human/Primate IL‑15 Biotinylated Antibody (Catalog # BAM247)
  • Standard: Recombinant Human IL-15 Protein (Catalog # 247-IL)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using MAB647 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 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. *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

P40933

Additional IL-15 Products

Product Documents for IL-15 Antibody (34505)

Certificate of Analysis

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Product Specific Notices for IL-15 Antibody (34505)

For research use only

Citations for IL-15 Antibody (34505)

Customer Reviews for IL-15 Antibody (34505) (2)

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Customer Images

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  • Human/Primate IL-15 Antibody
    Name: Katie Myers
    Application: ELISA
    Sample Tested: Serum
    Species: Primate
    Verified Customer | Posted 11/28/2022
    Used as an ELISA Capture IgG
    IL-15 Antibody (34505) MAB647
  • Human/Primate IL-15 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 11/20/2017
    This antibody was used as the solid phase molecule in an ELISA for IL15. The BAM247 antibody was used as the liquid phase molecule.
    IL-15 Antibody (34505) MAB647

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