Human/Rat IL-2 Antibody AF-502-NA: R&D Systems

Human/Rat IL-2 Antibody

(4 citations)
(2 Reviews)
  
  • Species Reactivity
    Human, Rat
  • 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.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant rat IL-2
    Ala21-Gln155
    Accession # P17108
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
    • Rat IL-2 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Human/Rat IL‑2 Antibody (Catalog # AF-502-NA)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Rat IL‑2 Biotinylated Antibody (Catalog # BAF502)
  • ELISA Standard
     
    Recombinant Rat IL-2 Protein (Catalog # 502-RL)
  • 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.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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 ug/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 (Catalog # 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.
Immunocytochemistry
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; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Immunohistochemistry
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; Catalog # 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. Recombinant Rat IL‑2 (Catalog # 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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 δ 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
  • Entrez Gene IDs:
    3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit)
  • Alternate Names:
    aldesleukin; IL2; IL-2; IL-2lymphokine; interleukin 2; interleukin-2; involved in regulation of T-cell clonal expansion; T cell growth factor; T-cell growth factor; TCGF
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Sample Type
  1. A Crucial Role of CXCL14 for Promoting Regulatory T Cells Activation in Stroke
    Authors: HT Lee, SP Liu, CH Lin, SW Lee, CY Hsu, HK Sytwu, CH Hsieh, WC Shyu
    Theranostics, 2017;7(4):855-875.
    Species: Rat
    Sample Type: In Vivo
    Application: Neut
  2. Antidiabetic Effect of Interleukin-1beta Antibody Therapy Through beta-Cell Protection in the Cohen Diabetes-Sensitive Rat.
    Authors: Aharon-Hananel G, Jorns A, Lenzen S, Raz I, Weksler-Zangen S
    Diabetes, 2015;64(5):1780-5.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  3. Differential control of T regulatory cell proliferation and suppressive activity by mature plasmacytoid versus conventional spleen dendritic cells.
    Authors: Ouabed A, Hubert FX, Chabannes D, Gautreau L, Heslan M, Josien R
    J. Immunol., 2008;180(9):5862-70.
    Species: Rat
    Sample Type: Whole Cells
    Application: Neut
  4. Overexpression of interleukin-13 induces minimal-change-like nephropathy in rats.
    Authors: Lai KW, Wei CL, Tan LK, Tan PH, Chiang GS, Lee CG, Jordan SC, Yap HK
    J. Am. Soc. Nephrol., 2007;18(5):1476-85.
    Species: Rat
    Sample Type: Serum
    Application: ELISA Development
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Average Rating: 4.5 (Based on 2 reviews)

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ELISA Human/Rat IL-2 Antibody AF-502-NA
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Excellent
 ELISA Anonymous 11/08/2017
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ELISA Human/Rat IL-2 Antibody AF-502-NA
ELISA: Human/Rat IL-2 Antibody [AF-502-NA]

Summary

ApplicationELISA
Sample TestedSerum

Other Experimental Details

Other Experimental DetailsThe rat polyclonal AF502 was paired with BAF502 to build an ELISA for the measurement of Rat IL-2 in serum samples.
ELISA Rat IL‑2 Antibody AF-502-NA
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Very Good
 ELISA Anonymous 10/26/2015
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ELISA Rat IL‑2 Antibody AF-502-NA
ELISA: Rat IL‑2 Antibody [AF-502-NA]

Summary

ApplicationELISA
Sample TestedCell culture supernatant

Other Experimental Details

Other Experimental DetailsELISA 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.
Specificity: Specific
Sensitivity: Sensitive
Buffer: Wash and blocking buffers
Dilution: 1.6ug/mL

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