Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, ELISA, Neutralization, Immunocytochemistry

Cited:

Neutralization, ELISA Capture

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

E. coli-derived recombinant human IL-2
Ala21-Thr153
Accession # P60568

Specificity

Detects human IL-2 in direct ELISAs and Western blots. In direct ELISAs, greater than 50% cross-reactivity with recombinant rabbit IL-2 is observed and less than 20% cross-reactivity with recombinant bovine IL-2, recombinant mouse IL-2 and recombinant rat IL-2 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 Human IL‑2 Antibody

Detection of Human IL-2 antibody by Western Blot.

Detection of Human IL‑2 by Western Blot.

Western blot shows lysates of monensin treated human peripheral blood mononuclear cells (PBMCs) with no additional treatment (-) or additionally treated (+) with 0.5 μg/mL calcium ionomycin (Iono) and 50 ng/mL PMA overnight. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for IL-2 at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

IL-2 antibody in Human PBMCs by Immunocytochemistry (ICC).

IL‑2 in Human PBMCs.

IL-2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PMA, ionomyocin, and monensin using Goat Anti-Human IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; (NL001) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Cell Proliferation Induced by IL‑2 and Neutralization by Human IL‑2 Antibody.

Cell Proliferation Induced by IL‑2 and Neutralization by Human IL‑2 Antibody.

Recombinant Human IL-2 (202-IL) stimulates proliferation in the CTLL-2 mouse cytotoxic T cell line in a dose-dependent manner (orange line) as measured by Resazurin (AR002). Proliferation elicited by Recombinant Human IL-2 (2 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA). The ND50 is typically ≤ 0.15 µg/mL.

Applications for Human IL‑2 Antibody

Application
Recommended Usage

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human IL‑2 Monoclonal Antibody (Catalog # MAB2021).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human IL-2 DuoSet ELISA Kit (Catalog # DY202) for convenient development of a sandwich ELISA or the Human IL-2 Quantikine ELISA Kit (Catalog # D2050) for a complete optimized ELISA.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells stimulated with PMA, ionomycin, and monensin

Western Blot

0.5 µg/mL
Sample:

Human peripheral blood mononuclear cells (PBMCs) treated with monensin, 0.5ug/mL calcium ionomycin and 50ng/mL PMA overnight

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 ≤0.15 µg/mL in the presence of 2 ng/mL Recombinant Human IL‑2.

Reviewed Applications

Read 2 reviews rated 5 using AF-202-NA in the following applications:

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.


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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-2

Interleukin-2 (IL-2) is a O-glycosylated, four alpha -helix bundle cytokine that has potent stimulatory activity for antigen-activated T cells. It is expressed by CD4+ and CD8+ T cells, gamma delta T cells, B cells, dendritic cells, and eosinophils (1 - 3). Mature human IL-2 shares 56% and 66% 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).

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.

Long Name

Interleukin 2

Alternate Names

Aldesleukin, IL2, Proleukin, TCGF

Entrez Gene IDs

3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit)

Gene Symbol

IL2

UniProt

Additional IL-2 Products

Product Documents for Human IL‑2 Antibody

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‑2 Antibody

For research use only

Citations for Human IL‑2 Antibody

Customer Reviews for Human IL‑2 Antibody (2)

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Showing  1 - 2 of 2 reviews Showing All
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  • Human IL-2 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum
    Species: Cynomolgus Monkey
    Verified Customer | Posted 12/12/2020
    Human IL‑2 Antibody AF-202-NA
  • Human IL-2 Antibody
    Name: Zoia Levashova
    Application: Meso Scale Discovery Assay
    Sample Tested: EDTA Plasma
    Species: Mouse
    Verified Customer | Posted 06/19/2018
    Used for detection of IL2 conjugated to the target-specific antibody. Used either as a capture or a detection antibody after labeling with biotin or sulfo-tag, respectively, according to the manufacturer’s protocol (Meso Scale Diagnostics LLC). Detection range of IL-2 conjugates was 10-225,000 pg/ml for anti-IL2-Biotin/anti-Fc gamma-Sulfo-Tag (A) and 50-225,000 pg/ml for anti-Fc gamma-Biotin/anti-IL2-Sulfo-Tag capture/detection combination (B).
    Human IL‑2 Antibody AF-202-NA

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Protocols

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