Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

Neutralization, Intracellular Staining by Flow Cytometry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 5334R
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Product Specifications

Immunogen

E. coli-derived recombinant human IL-2
Ala21-Thr153
Accession # NP_000577

Specificity

Detects human IL-2 in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human IL‑2 Antibody

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) treated with calcium ionomycin and PMA using Mouse Anti-Human IL-2 Monoclonal Antibody (Catalog # MAB202R) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. 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 (Catalog # 202-IL) stimulates proliferation in the CTLL-2 mouse cytotoxic T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL-2 (2 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-2 Monoclonal Antibody (Catalog # MAB202). The ND50 is typically 0.015-0.03 µg/mL.

Detection of IL-2 in Human PBMCs by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs) (A) treated with Cell Activation Cocktail 500x (5476) for 5 hours or (B) resting were stained with Mouse Anti-Human IL-2 Monoclonal Antibody (Catalog # MAB202R) followed by Goat anti-Mouse IgG APC-conjugated Secondary Antibody (F0101B) and Mouse anti-Human CD3 PE-conjugated Monoclonal Antibody (FAB100P). Quadrant markers were set based on Mouse IgG1 isotype control antibody (MAB002). To facilitate intracellular staining, cells were fixed and permeabilized using FlowX FoxP3/Transcription Factor Fixation & Perm Buffer Kit (FC012). Staining was performed using our Staining Intracellular Molecules protocol.

Detection of Human IL-2 by Flow Cytometry

Detection of Human IL-2 by Flow Cytometry

IL-2 induces IL-32 but TCR activation is required for IL-32 secretion. (A) IL-32 expression in CD3+ T from healthy donors (HDs) after 24 h of resting (unstimulated) either alone or in the presence of IFN-gamma & low (100 IU/ml) or high (3,000 IU/ml) amounts of IL-2 & after 24 h of either CD3 or CD3/CD28 stimulation determined by ICFC analysis. Representative data from n=2 independent experiments. (B–D) WB analyses of IL-32 expression in cell lysates & IL-32 secretion into the cell culture supernatants (enriched via Amicon Ultracentrifugal filters) by HD CD3+ T after treatment for 48 h with 500 or 3,000 IU/ml IL-2 alone (low or high IL-2, respectively) or treatment with high IL-2 for 48 h with subsequent CD3/CD28 stimulation for 72 (h) (B) Representative blot of lysates & supernatants (15 µg) from n=2 HDs (#1, #2) next to rIL-32 beta (4 ng, MW: 23.1 kDa) & rIL-32 gamma (4 ng, MW: 28.1 kDa). As a loading control, the housekeeping gene GAPDH (MW: 37 kDa) was detected on same blot. (C) Cumulative data of quantified IL-32 beta expression in cell lysates normalized to GAPDH for n=2–5 HDs, mean+SD, dots depict data from individual experiments, Student’s paired t-test, *p<0.05. (D) Cumulative data of quantified IL-32 beta secretion into culture supernatants displayed by the Area Under the Curve (AUC) for n=2–5 HDs, mean+SD. (E, F) IL-32 expression in HD T after resting (Unstim) or CD3/CD28 stimulation alone (Stim) or together with either an alpha IL-2 neutralizing antibody (Stim + alpha IL-2) or the respective isotype control (Stim + Isotype) for 24 (h) (E) Representative dot plots depicting IL32+CD25+ among live single T stimulated in the presence of the isotype or the alpha IL-2 antibody. (F) Cumulative data showing the frequency of IL-32+CD25+ among live single T, n=2 HDs, Student’s unpaired t-test, *p<0.05. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39211051), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human IL‑2 Antibody

Application
Recommended Usage

Immunocytochemistry

5-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with calcium ionomycin and PMA

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood mononuclear cells (PBMCs) treated with Cell Activation Cocktail 500x (Catalog # 5476) for 5 hours,  fixed and permeabilized using FlowX FoxP3/Transcription Factor Fixation & Perm Buffer Kit (Catalog # FC012)

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 0.015-0.03 µg/mL in the presence of 2 ng/mL Recombinant Human IL‑2.

Flow Cytometry Panel Builder

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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 cell culture supernatant

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. *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 cytokine that stimulates the growth and differentiation of B cells, T cells, NK cells, and monocyte/macrophages. It functions through the heterotrimeric IL-2 receptor comprising alpha, beta, and gamma chains.

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

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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

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Protocols

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