IL-2 Antibody [Unconjugated]

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.

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 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 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.

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.

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.

• Reactivity: Hu, Rt
• Applications: WB, IHC, B/N, ELISA(Cap), ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

IL-2 Antibody [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: IL2
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • ELISA Capture (Matched Antibody Pair) 0.2-0.8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • ELISA Standard (Matched Pair)
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 3-15 ug/mL
  • Neutralization 0.15-0.75 ug/mL
  • Western Blot 2 ug/mL
• Application Notes: ELISA Capture: Rat IL-2 Antibody (Catalog # AF-502-NA)
  ELISA Detection: Rat IL-2 Biotinylated Antibody (Catalog # BAF502)
  Standard: Recombinant Rat IL-2 (Catalog # 502-RL)

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for IL-2 Antibody [Unconjugated]

• Aldesleukin
• IL2
• IL-2
• IL-2lymphokine
• interleukin 2
• interleukin-2
• involved in regulation of T-cell clonal expansion
• Proleukin
• T cell growth factor
• T-cell growth factor
• TCGF

Background

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).

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.

Show all background references

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IL-2 Antibody (AF-502-NA)(5)

Publications using AF-502-NA

• W Cao, F Fang, T Gould, X Li, C Kim, C Gustafson, S Lambert, CM Weyand, JJ Goronzy Ecto-NTPDase CD39 is a negative checkpoint that inhibits follicular helper cell generation J. Clin. Invest., 2020-07-01;0(0):. 2020-07-01 [PMID: 32452837] (Neutralization, Human)
• HT Lee, SP Liu, CH Lin, SW Lee, CY Hsu, HK Sytwu, CH Hsieh, WC Shyu A Crucial Role of CXCL14 for Promoting Regulatory T Cells Activation in Stroke Theranostics, 2017-02-08;7(4):855-875. 2017-02-08 [PMID: 28382159] (Neutralization, Rat)
• Aharon-Hananel G, Jorns A, Lenzen S, Raz I, Weksler-Zangen S Antidiabetic Effect of Interleukin-1beta Antibody Therapy Through beta-Cell Protection in the Cohen Diabetes-Sensitive Rat. Diabetes, 2014-12-08;64(5):1780-5. 2014-12-08 [PMID: 25488902] (IHC-P, Rat)
• Ouabed A, Hubert FX, Chabannes D, Gautreau L, Heslan M, Josien R Differential control of T regulatory cell proliferation and suppressive activity by mature plasmacytoid versus conventional spleen dendritic cells. J. Immunol., 2008-05-01;180(9):5862-70. 2008-05-01 [PMID: 18424705] (Neutralization, Rat)
• Lai KW, Wei CL, Tan LK, Tan PH, Chiang GS, Lee CG, Jordan SC, Yap HK Overexpression of interleukin-13 induces minimal-change-like nephropathy in rats. J. Am. Soc. Nephrol., 2007-04-11;18(5):1476-85. 2007-04-11 [PMID: 17429054] (ELISA Development, Rat)

Bioinformatics

• Gene Symbol: IL2
• Uniprot:
  • P17108()