Human IL-2 Antibody (5334R)

Name: Human IL-2 Antibody (5334R)
Brand: R&D Systems
SKU: MAB202R
Price: 489 USD
Availability: InStock

R&D Systems | Catalog # MAB202R

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Product Documents

Recombinant Monoclonal Antibody.

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Product Specifications
Scientific Data
Applications
Flow Cytometry
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

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

View all IL-2 Primary Antibodies »

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 (5334R)

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.

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

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.

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Applications for Human IL-2 Antibody (5334R)

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.

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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.

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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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)