Human/Rat IL-2 Antibody Summary
Accession # P17108
Rat IL-2 Sandwich Immunoassay
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.
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 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 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. 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.
Preparation and Storage
- 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.
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).
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Citations for Human/Rat IL-2 Antibody
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.
Citations: Showing 1 - 5
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Ecto-NTPDase CD39 is a negative checkpoint that inhibits follicular helper cell generation
Authors: W Cao, F Fang, T Gould, X Li, C Kim, C Gustafson, S Lambert, CM Weyand, JJ Goronzy
J. Clin. Invest., 2020;0(0):.
Sample Types: Whole Cells
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
Sample Types: In Vivo
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
Sample Types: Whole Tissue
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.
Sample Types: Whole Cells
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.
Sample Types: Serum
Applications: ELISA Development
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The rat polyclonal AF502 was paired with BAF502 to build an ELISA for the measurement of Rat IL-2 in serum samples.
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.
Buffer: Wash and blocking buffers