IL-2 Antibody

R&D Systems | Catalog # AF-502-NA

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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Rat

Cited:

Human, Rat

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Neutralization, Immunocytochemistry, ELISA Development

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all IL-2 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant rat IL-2
Ala21-Gln155
Accession # P17108

Specificity

Detects rat IL-2 in ELISAs. Detects human and rat IL-2 in Western blots. In sandwich immunoassays, less than 0.2% cross-reactivity with recombinant mouse 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 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 2 µg/mL of Goat Anti-Human/Rat IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-502-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 Rat Splenocytes by Immunocytochemistry (ICC).

IL‑2 in Rat Splenocytes.

IL-2 was detected in immersion fixed rat splenocytes stimulated with calcium ionomycin and PMA using Goat Anti-Human/Rat IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-502-NA) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
IL-2 antibody in Rat Spleen by Immunohistochemistry (IHC-Fr).

IL‑2 in Rat Spleen.

IL-2 was detected in immersion fixed frozen sections of rat spleen using Goat Anti-Human/Rat IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-502-NA) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Cell Proliferation Induced by IL‑2 and Neutralization by Rat IL‑2 Antibody.

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

Recombinant Rat IL-2 (502-RL) stimulates proliferation in the CTLL-2 mouse cytotoxic T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat IL-2 (2 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Rat IL-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-502-NA). The ND50 is typically 0.15-0.75 µg/mL.

Detection of IL‑2 in Human Tonsil.

IL‑2 was detected in immersion fixed paraffin-embedded sections of Human Tonsil using Goat Anti-Human/Rat IL‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-502-NA) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Rat IL-2 ELISA Standard Curve

Rat IL-2 ELISA Standard Curve

Recombinant Rat IL‑2 (Catalog # 502-RL) was serially diluted and captured by Goat Anti-Human/Rat IL‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-502-NA) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human/Rat IL‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-502-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Rat IL-2 ELISA Standard Curve

Rat IL-2 ELISA Standard Curve

Recombinant Rat IL‑2 (Catalog # 502-RL) was serially diluted and captured by Goat Anti-Human/Rat IL‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-502-NA) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human/Rat IL‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-502-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for IL-2 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed rat splenocytes stimulated with calcium ionomycin and PMA

Immunohistochemistry

3-15 µg/mL
Sample: Immersion fixed frozen sections of Rat Spleen and Human Tonsil

Western Blot

2 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 0.15-0.75 µg/mL in the presence of 2 ng/mL Recombinant Rat IL‑2.

Rat IL-2 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Rat IL‑2 Biotinylated Antibody (Catalog # BAF502)
  • Standard: Recombinant Rat IL-2 Protein (Catalog # 502-RL)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 4.5 using AF-502-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.

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 rat IL-2 shares 66% and 73% amino acid sequence identity with human and mouse IL-2, respectively. The receptor for IL-2 consists of three subunits that are present on the cell surface in varying preformed complexes (4-6). 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 (7). IL-2 plays a central role in the expansion and maintenance of regulatory T cells, although it inhibits the development of Th17 polarized cells (8-10). Thus, IL-2 may be a key cytokine in the natural suppression of autoimmunity (11, 12).

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. McKnight, A. et al. (1989) Immunogenetics 30:145.
  4. Liparoto, S.F. et al. (2002) Biochemistry 41:2543.
  5. Wang, X. et al. (2005) Science 310:1159.
  6. Bodnar, A. et al. (2008) Immunol. Lett. 116:117.
  7. Jaleco, S. et al. (2003) J. Immunol. 171:61.
  8. Malek, T.R. (2003) J. Leukoc. Biol. 74:961.
  9. Laurence, A. et al. (2007) Immunity 26:371.
  10. Kryczek, I. et al. (2007) J. Immunol. 178:6730.
  11. Afzali, B. et al. (2007) Clin. Exp. Immunol. 148:32.
  12. 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

P17108

Additional IL-2 Products

Product Documents for IL-2 Antibody

Certificate of Analysis

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Product Specific Notices for IL-2 Antibody

For research use only

Customer Reviews for IL-2 Antibody (2)

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  • Human/Rat IL-2 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum
    Species: Rat
    Verified Customer | Posted 11/08/2017
    The rat polyclonal AF502 was paired with BAF502 to build an ELISA for the measurement of Rat IL-2 in serum samples.
    IL-2 Antibody AF-502-NA
  • Rat IL‑2 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 10/26/2015
    ELISA plates were coated by incubating the diluted capture antibody (AF-502-NA in PBS at 1.6ug/mL) overnight at 4C. To detect the captured IL2 from the supernatants being tested, the detection antibody BAF502 was used at 400ng/mL followed by avidin-HRP. <br />Specificity: Specific<br />Sensitivity: Sensitive<br />Buffer: Wash and blocking buffers<br />Dilution: 1.6ug/mL
    IL-2 Antibody AF-502-NA

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