Feline IL-2 Antibody

Catalog #: AF1890
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Catalog #	Availability	Size / Price	Qty
AF1890
AF1890-SP

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Feline IL-2 Antibody Summary

Species Reactivity

Feline

Specificity

Detects feline IL‑2 in ELISAs and Western blots. In sandwich immunoassays, approximately 3% cross-reactivity with recombinant canine IL-2 is observed, approximately 1% cross reactivity with recombinant human IL-2 and recombinant porcine IL-2 is observed, and less than 0.2% cross‑reactivity with recombinant mouse IL-2, recombinant equine IL-2, recombinant bovine IL-2, recombinant rat IL-2, and recombinant cotton rat IL-2 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant feline IL‑2
Ala21-Thr154 (Cys146Ser)
Accession # Q07885

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.

Endotoxin Level

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

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Feline IL‑2 (Cys146Ser) (Catalog # 1890-FL)

Immunocytochemistry

5-15 µg/mL

See below

Feline IL-2 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

0.2-0.8 µg/mL

Use in combination with:

Detection Reagent: Feline IL‑2 Biotinylated Antibody (Catalog # BAF1890)

Standard: Recombinant Feline IL-2 (Cys146Ser) Protein (Catalog # 1890-FL)

Neutralization

Measured by its ability to neutralize IL‑2-induced proliferation in the CTLL‑2 mouse cytotoxic T cell line. Gearing, A. J. H. and C. B. Bird (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M. J. et al. (eds): IRL Press. 276. The Neutralization Dose (ND₅₀) is typically 0.03-0.15 µg/mL in the presence of 0.2 ng/mL Recombinant Feline IL‑2 (Cys146Ser).

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.

Scientific Data

Immunocytochemistry

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IL‑2 in Feline PBMCs. IL-2 was detected in immersion fixed feline peripheral blood mononuclear cells (PBMCs) stimulated with PMA and calcium ionomycin using Goat Anti-Feline IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1890) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Neutralization

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Cell Proliferation Induced by IL‑2 and Neutralization by Feline IL‑2 Antibody. Recombinant Feline IL-2 (Cys146Ser) (Catalog # 1890-FL) induces cell 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 Feline IL-2 (Cys146Ser) (0.2 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Feline IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1890). The ND₅₀ is typically 0.03-0.15 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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-2

Interleukin-2 (IL-2) is a secreted, single chain alpha -helical polypeptide that has potent stimulatory activity for antigen-activated T cells. The feline IL-2 gene encodes a 154 amino acid (aa) precursor protein with a 20 aa signal peptide plus a 134 aa mature segment. There are suggestions that the mature protein may be
O‑glycosylated. At the aa sequence level, mature feline IL-2 is 78%, 82%, 60%, 64%, 62%, 75%, 62%, and 76% identical to mature human, canine, mouse, rat, cotton rat, porcine, goat, and equine IL-2, respectively. Mammalian cells known to express IL-2 include CD4⁺ and CD8⁺ T cells, visceral smooth muscle cells, eosinophils, gamma δ T cells, B cells and dendritic cells. The biological activity of IL-2 is mediated by IL-2 receptor complexes consisting of three distinct subunits ( alpha, beta, gamma ) in two combinations. The high-affinity signaling IL-2 receptor complex is a heterotrimer of the IL-2 receptor alpha, beta, gamma subunits. The intermediate signaling complex is a heterodimer of the IL-2 R beta and gamma subunits. The non-ligand binding gamma subunit, referred to as the common gamma subunit ( gamma _c), is also a subunit of the receptor complexes of IL-4, IL-7, IL-9 and IL-15. Functionally, IL-2 is best known for its autocrine and paracrine activity on T cells. On naïve CD8⁺ T cells, high IL-2 levels can induce cell proliferation with a bias towards cytotoxicity. In the presence of low levels of IL-2, CD8⁺ T cells preferentially undergo apoptosis with a bias towards cytokine secretion. IL-2 also seems to play a central role in the expansion and maintenance of CD4⁺ CD25⁺ regulatory T cells. This indicates IL-2 may be a key cytokine in the natural suppression of autoimmunity (1-9).

References

Cozzi, P.J. et al. (1993) Biochem. Biophys. Res. Commun. 194:1038.
Smith, K.A. (1992) Curr. Opin. Immunol. 4:271.
Conradt, H.S. et al. (1989) J. Biol. Chem. 264:17368.
Matesanz, F. et al. (1993) Immunogenetics 38:300.
Ellery, J.M. and P.J. Nicholls (2002) Cytokine Growth Factor Rev. 13:27.
Liparoto, S.F. et al. (2002) Biochemistry 41:2543.
Lin, J-X. and W.J. Leonard (1997) Cytokine Growth Factor Rev. 8:313.
Sad, S. and L. Krishnan (1999) J. Immunol. 163:2443.
Malek, T.R. (2003) J. Leukoc. Biol. 74:961.

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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Citation for Feline IL-2 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

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