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Human IL-2 Antibody (5334)

R&D Systems | Catalog # MAB202

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Xenograft

Applications

Validated:

Neutralization, Intracellular Staining by Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Neutralization, Flow Cytometry, Immunocytochemistry, Bioassay, ELISA Detection, In vivo assay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 5334
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. Does not cross-react with recombinant IL-2 from mouse, rat, pig, or cotton rat.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Endotoxin Level

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

Scientific Data Images for Human IL-2 Antibody (5334)

Detection of IL‑2 in PBMC's treated with PMA and Ca2+ ionomycin vs untreated PBMC's by Flow Cytometry

PBMC's treated with PMA (50ng/mL) and Ca2+ ionomycin (200ng/mL) overnight (A) vs untreated PBMC's (B) were stained with Mouse Anti-Human IL‑2 Monoclonal Antibody (Catalog # mab202) and Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A) followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
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.

Human IL 2 Antibody (Catalog # MAB202) neutralizes IL-2-induced proliferation in the CTLL-2 mouse cytotoxic T cell line. The Neuralization Dose (ND50) is typically 5.00-60.0 ng/mL.
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 # MAB202) at 8 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.
Detection of Human IL-2 by Flow Cytometry

Detection of Human IL-2 by Flow Cytometry

SLE CD4+ T cells are poised to activate IL-2 signaling during Treg differentiation. (A) Naïve CD4+ T cells were isolated from a systemic lupus erythematosus (SLE) patient & matched healthy control (HC) subject, & cultured for 3 days in the presence of anti-CD3/CD28 & TGF-beta (5 ng/ml) with IL-2 (50 IU/ml) or anti-IL-2 (100 or 1,000 ng/ml). The frequency of CD4+CD25+FOXP3+ cells was determined by flow cytometry. Numbers below the plots represent the frequency of CD4+CD25+FOXP3+ Tregs. The dot plots on the left end represent isotype control staining. (B) CD4+ T cells isolated from matched SLE & HC subjects were cultured for 3 days in the presence of anti-CD3/CD28 & TGF-beta (20 ng/ml) with or without IL-2 (100 IU/ml) or anti-IL-2 (100 ng/ml). Total STAT5 & its phosphorylation at tyrosine 694 were detected by immunoblotting. Representative immunoblot staining (left panel). The signal intensity of phospho-STAT5 & total STAT5 was normalized to that of actin. The normalized pSTAT5 signal intensity (middle panel) & the ratio of normalized pSTAT5 signal intensity over normalized STAT5 signal intensity (right panel) from 3 pairs of matched HC & SLE subjects. (C) Untouched T cells from matched SLE & HC subjects were cultured for 3 days without anti-CD3/CD28 stimulation. Expression of CD25 & FOXP3 in CD4+ cells were determined by flow cytometry. Representative flow cytometry dot plots are shown (left panel). Cumulative data of frequency of CD4+CD25+FOXP3+ & CD4+CD25+ cells, mean fluorescence intensity (MFI) of CD25 expression in CD4+ T cells, & the proportion of CD4+CD25+FOXP3+ cells among CD4+CD25+ cells from 17 pairs of matched SLE & HC subjects (right panel). Data were analyzed by a paired two-tailed t-test (*p<0.05, **p<0.01, ****p<0.0001). Image collected & cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fimmu.2021.635531/full), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-2 by Western Blot

Detection of Human IL-2 by Western Blot

SLE CD4+ T cells are poised to activate IL-2 signaling during Treg differentiation. (A) Naïve CD4+ T cells were isolated from a systemic lupus erythematosus (SLE) patient & matched healthy control (HC) subject, & cultured for 3 days in the presence of anti-CD3/CD28 & TGF-beta (5 ng/ml) with IL-2 (50 IU/ml) or anti-IL-2 (100 or 1,000 ng/ml). The frequency of CD4+CD25+FOXP3+ cells was determined by flow cytometry. Numbers below the plots represent the frequency of CD4+CD25+FOXP3+ Tregs. The dot plots on the left end represent isotype control staining. (B) CD4+ T cells isolated from matched SLE & HC subjects were cultured for 3 days in the presence of anti-CD3/CD28 & TGF-beta (20 ng/ml) with or without IL-2 (100 IU/ml) or anti-IL-2 (100 ng/ml). Total STAT5 & its phosphorylation at tyrosine 694 were detected by immunoblotting. Representative immunoblot staining (left panel). The signal intensity of phospho-STAT5 & total STAT5 was normalized to that of actin. The normalized pSTAT5 signal intensity (middle panel) & the ratio of normalized pSTAT5 signal intensity over normalized STAT5 signal intensity (right panel) from 3 pairs of matched HC & SLE subjects. (C) Untouched T cells from matched SLE & HC subjects were cultured for 3 days without anti-CD3/CD28 stimulation. Expression of CD25 & FOXP3 in CD4+ cells were determined by flow cytometry. Representative flow cytometry dot plots are shown (left panel). Cumulative data of frequency of CD4+CD25+FOXP3+ & CD4+CD25+ cells, mean fluorescence intensity (MFI) of CD25 expression in CD4+ T cells, & the proportion of CD4+CD25+FOXP3+ cells among CD4+CD25+ cells from 17 pairs of matched SLE & HC subjects (right panel). Data were analyzed by a paired two-tailed t-test (*p<0.05, **p<0.01, ****p<0.0001). Image collected & cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fimmu.2021.635531/full), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-2 by Flow Cytometry

Detection of Human IL-2 by Flow Cytometry

IL-2 expands IL-13+IFN-gamma + CD8+ T cells in systemic lupus erythematosus (SLE). (A) CD4+ and CD8+ T cells from matched SLE and health control (HC) subjects were cultured as described in Figure 2, and IL-13 and IFN-gamma expression was determined by flow cytometry. (B) Cumulative data of mean fluorescence intensity (MFI) of IFN-gamma and the frequency of IFN-gamma + and IL-13+IFN-gamma + cells. Statistical analysis was made by two-way ANOVA followed by Bonferroni’s correction for multiple comparisons (*p<0.05, **p<0.01, ****p<0.0001). Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fimmu.2021.635531/full), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-2 by Western Blot

Detection of Human IL-2 by Western Blot

IL-2 induces STAT6 phosphorylation and GATA3 expression in systemic lupus erythematosus (SLE) CD8+ T cells. (A) CD8+ T cells from matched SLE and health control (HC) subjects were cultured as described in Figure 2. Expression of GATA-3 and phosphorylation of STAT5 at tyrosine 694 and STAT6 at tyrosine 641 were detected by immunoblotting. Representative immunoblot staining was presented. Lo and Hi concentrations of anti-IL-2 denote 100 and 1,000 ng/ml, respectively. (B) The signal intensity of phospho-STAT5, phospho-STAT6, and GATA-3 were normalized to that of actin. Cumulative data from 9 pairs of matched HC and SLE subjects. Data were analyzed by a two-tailed t-test (*p<0.05, **p<0.01, ***p<0.001). (C) Pearson’s and Spearman’s correlation analyses were performed to determine the association between the expression of cytokines (IL-13, IL-5, and IFN-gamma ) and transcription factors (phospho-STAT5, phospho-STAT6, and GATA-3). The blue and red plots represent data from HC and SLE patients, respectively. Spearman correlation coefficient was presented for the association between IL-13 and phospho-STAT5, IL-5 and phospho-STAT-5, and IFN-gamma and GATA-3. Pearson correlation coefficient was presented for the remainder of associations. Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fimmu.2021.635531/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human IL-2 Antibody (5334)

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells treated with calcium ionomycin and PMA

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood mononuclear cells treated with PMA and Ca2+ ionomycin, fixed with paraformaldehyde, and permeabilized with saponin

Neutralization

Measured by its ability to neutralize IL-2-induced proliferation in the CTLL-2 mouse cytotoxic T cell line. Gearing, A.H.H. and C.B. Bird (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 276. The Neuralization Dose (ND50) is typically 5.00-60.0 ng/mL in the presence of 2 ng/mL Recombinant Human IL-2.

Reviewed Applications

Read 2 reviews rated 4.5 using MAB202 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma 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. See Certificate of Analysis for details.
*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

NP_000577

Additional IL-2 Products

Product Documents for Human IL-2 Antibody (5334)

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.

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Product Specific Notices for Human IL-2 Antibody (5334)

For research use only

Citations for Human IL-2 Antibody (5334)

Customer Reviews for Human IL-2 Antibody (5334) (2)

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Customer Images

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  • Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: BUD-8 human fibroblast cell line
    Species: Human
    Verified Customer | Posted 11/16/2020
  • Human IL-2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Serum
    Species: Human
    Verified Customer | Posted 12/12/2019
    Human IL-2 Antibody (5334) MAB202

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Protocols

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FAQs for Human IL-2 Antibody (5334)

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  • Q: Can Catalog # MAB202 (anti-Human IL-2 antibody) be used for neutralization of mouse IL-2?

    A: No. MAB202 is not cross-reactive to mouse IL-2 and so cannot be used for neutralization of mouse IL-2.

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