Human IL-2 Alexa Fluor® 647-conjugated Antibody

Catalog #: IC103561R Datasheet / COA / SDS
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IC103561R-100UG
R&D Systems Antibodies
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Human IL-2 Alexa Fluor® 647-conjugated Antibody Summary

Species Reactivity
Human
Specificity
Detects human IL-2 in direct ELISA.
Source
Monoclonal Mouse IgG1 Clone # 1019308
Purification
Protein A or G purified from cell culture supernatant
Immunogen
E. coli-derived human IL-2
Ala21-Thr153
Accession # P60568
Formulation
Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 647 (Excitation= 650 nm, Emission= 668 nm)

Applications

Recommended Concentration
Sample
Flow Cytometry
Titration recommended for optimal concentration with starting range of 0.1-1 µg/1 million cells. Sample used for this experiment was PBMC stimulated vs naïve PBMC lymphocytes.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

Background: IL-2

Recombinant Interleukin-2 (IL-2) is expressed in E. coli and has been engineered to contain the serine for cysteine substitution found in Proleukin® (aldesleukin). Recombinant IL-2 is widely used in cell culture for the expansion of T cells. IL-2 is expressed by CD4+ and CD8+ T cells, gamma δ T cells, B cells, dendritic cells, and eosinophils (1-3). Mature human IL-2 shares 56% and 66% amino acid (aa) sequence identity with mouse and rat IL-2, respectively. Human and mouse IL-2 exhibit cross-species activity (4). The receptor for IL-2 consists of three subunits that are present on the cell surface in varying preformed complexes (5-7). The 55 kDa IL-2 R alpha is specific for IL-2 and binds with low affinity. The 75 kDa IL-2 R beta, which is also a component of the IL-15 receptor, binds IL-2 with intermediate affinity. The 64 kDa common gamma chain gamma c/IL-2 R gamma, which is shared with the receptors for IL-4, -7, -9, -15, and -21, does not independently interact with IL-2. Upon ligand binding, signal transduction is performed by both IL-2 R beta and gamma c.

IL-2 is best known for its autocrine and paracrine activity on T cells. It drives resting T cells to proliferate and induces IL-2 and IL-2 R alpha synthesis (1, 2). It contributes to T cell homeostasis by promoting the Fas-induced death of naïve CD4+ T cells but not activated CD4+ memory lymphocytes (8). IL-2 plays a central role in the expansion and maintenance of regulatory T cells, although it inhibits the development of Th17 polarized cells (9-11). Thus, IL-2 may be a key cytokine in the natural suppression of autoimmunity (12, 13).

IL-2 expression and concentration can have either immunostimulatory effects at high doses or immunosuppressive effects at low doses due to its preferential binding to different receptor subunits expressed by various immune cell types. This has led to the generation of recombinant IL-2 variants aimed at modifying IL-2 receptor binding for increased antitumor efficacy (14, 15). These variants are typically used in combination with immune checkpoint inhibitors instead of as a monotherapy (14). IL-2 can be genetically engineered to express in NK cells for CAR T cell therapies, and in combination with other cytokines like IL-15, can increase cell viability and proliferation (16). In addition to adoptive cell transfer and checkpoint blockade inhibitors, cancer vaccines that boost immune responses have been combined with IL-2 treatment with promising results in recent studies (15).

In cell culture, IL-2 is a frequently used cytokine for the proliferation, differentiation, and increased antibody secretion of B cells as they transform into plasma cells in vitro (17). IL-2 is also a classically used cytokine for the expansion of NK cells, early differentiated T cells and effector memory Treg cells for adoptive cell transfer cancer immunotherapy (16, 18). GMP IL-2 is a commonly used supplement for the expansion of these cell types for cellular therapies.

References
  1. Ma, A. et al. (2006) Annu. Rev. Immunol. 24:657.
  2. Gaffen, S.L. and K.D. Liu (2004) Cytokine 28:109.
  3. Taniguchi, T. et al. (1983) Nature 302:305.
  4. Mosmann, T.R. et al. (1987) J. Immunol. 138:1813.
  5. Liparoto, S.F. et al. (2002) Biochemistry 41:2543.
  6. Wang, X. et al. (2005) Science 310:1159.
  7. Bodnar, A. et al. (2008) Immunol. Lett. 116:117.
  8. Jaleco, S. et al. (2003) J. Immunol. 171:61.
  9. Malek, T.R. (2003) J. Leukoc. Biol. 74:961.
  10. Laurence, A. et al. (2007) Immunity 26:371.
  11. Kryczek, I. et al. (2007) J. Immunol. 178:6730.
  12. Afzali, B. et al. (2007) Clin. Exp. Immunol. 148:32.
  13. Fehervari, Z.et al. (2006) Trends Immunol. 27:109.
  14. Xue, D. et al. (2021) Antibody Therapeutics. 4(2): 123-133.
  15. Wolfarth, A.A. et al. (2022) Immune Netw. 22(1): e5.
  16. Koehl, U. et al. (2015) Oncoimmunology. 5(4).
  17. Marsman, C. et al. (2022) Front. In Immunol. 13(815449).
  18. Chamucero-Millares, J.A. et al. (2021) Cellular Immunology. 360 (104257).
Long Name
Interleukin 2
Entrez Gene IDs
3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit)
Alternate Names
Aldesleukin; IL2; IL-2; IL-2lymphokine; interleukin 2; interleukin-2; involved in regulation of T-cell clonal expansion; Proleukin; T cell growth factor; T-cell growth factor; TCGF

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Product Specific Notices


This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

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