Mouse CD25/IL-2 R alpha Antibody

  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse CD25/IL-2 R alpha in ELISAs and Western blots. In sandwich ELISAs, less than 0.1% cross-reactivity with recombinant human CD25/IL-2 R alpha is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse CD25/IL-2 R alpha
    Glu22-Lys236
    Accession # Q544I2
  • 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
    0.1 µg/mL
    Recombinant Mouse CD25/IL‑2 R alpha (Catalog # 2438-RM)
  • Immunohistochemistry
    5-15 µg/mL
    See below
    • Mouse CD25/IL-2 R alpha Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Mouse CD25/IL-2 R alpha Antibody (Catalog # AF2438)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Mouse CD25/IL-2 R alpha Biotinylated Antibody (Catalog # BAF2438)
  • ELISA Standard
     
    Recombinant Mouse CD25/IL-2 R alpha Protein, CF (Catalog # 2438-RM)
  • Neutralization
    Measured by its ability to neutralize IL‑2-induced proliferation in the CTLL‑2 mouse cytotoxic T cell line. The Neutralization Dose (ND50) is typically 5-20 µg/mL in the presence of 2 ng/mL Recombinant Mouse 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

Cell Proliferation Induced by IL‑2 and Neutralization by Mouse CD25IL‑2 R alpha Antibody. Recombinant Mouse IL‑2 (Catalog # 402-ML) stimulates proliferation in the CTLL‑2 mouse cytotoxic T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Mouse IL‑2 (2 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse CD25/IL‑2 R alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2438). The ND50 is typically 5-20 µg/mL.

Immunohistochemistry
CD25/IL‑2 R alpha in Mouse Spleen.

CD25/IL‑2 R alpha was detected in immersion fixed frozen sections of mouse spleen using 5 µg/mL Goat Anti-Mouse CD25/IL‑2 R alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2438) overnight at 4 °C. Tissue was stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained (green). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

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: CD25/IL-2 R alpha

IL-2 receptor alpha (IL-2 R alpha ), also known as CD25, is a 55 kDa type I membrane glycoprotein that belongs to the family of cytokine receptors that utilize the common gamma chain subunit ( gamma c). IL-2 R alpha is primarily expressed on activated T cells and on regulatory T cells (Treg) (1-3). The mouse IL-2 R alpha cDNA encodes a 268 amino acid (aa) precursor that includes a 21 aa signal peptide, a 215 aa extracellular domain (ECD) with two Sushi domains, a 21 aa transmembrane segment, and an 11 aa cytoplasmic domain (4, 5). Within the ECD, mouse IL-2 R alpha shares 81% and 58% aa sequence identity with rat and human IL-2 R alpha, respectively. It shares approximately 15% aa sequence identity with IL-4, -7, -9, -15, and -21 receptor subunits that also complex with gamma c. IL-2 R beta (CD122) and gamma c (IL-2 R gamma /CD132) dimerize to form a constitutively expressed intermediate affinity IL-2 receptor (6, 7). By itself, IL-2 R alpha binds IL-2 with low affinity. It associates with IL-2 R beta and gamma c to generate a ternary high affinity IL-2 receptor complex (8). A soluble form of IL-2 R alpha can be generated by proteolytic cleavage of the cell surface receptor, rendering the T cell unresponsive to IL-2 (9, 10). Increased serum levels of soluble IL-2 R alpha are found in some cancers and immune disorders (11). IL-2 R alpha is required for activation induced cell death (AICD) of naive T cells, a mechanism responsible for deleting autoreactive T cell clones (12, 13). IL-2 R alpha is also required for the development of CD4+CD25+ Treg which suppress autoreactive CD4+ T cells, thereby contributing to peripheral T cell homeostasis (12-14).

  • References:
    1. Minami, Y. et al. (1993) Annu. Rev. Immunol. 11:245.
    2. Kovanen, P.E. and W.J. Leonard (2004) Immunol. Rev. 202:67.
    3. Bluestone, J.A. and Q. Tang (2005) Curr. Opin. Immunol. 17:638.
    4. Miller, J. et al. (1985) J. Immunol. 134:4212.
    5. Shimuzu, A. et al. (1985) Nucleic Acids Res. 13:1505.
    6. Hatakeyama, M. et al. (1989) Science 244:551.
    7. Takeshita, T. et al. (1992) Science 257:379.
    8. Wang, X. et al. (2005) Science 310:1159.
    9. Wagner, D.K. et al. (1986) J. Immunol. 137:592.
    10. Schulz, O. et al. (1998) J. Exp. Med. 187:271.
    11. Witkowska, A.M. (2005) Mediat. Inflamm. 2005:121.
    12. Willerford, D.M. et al. (1995) Immunity 3:521.
    13. Van Parijs, L. et al. (1997) J. Immunol. 158:3738.
    14. Almeida, A.R.M. et al. (2002) J. Immunol. 169:4850.
  • Long Name:
    Interleukin 2 Receptor alpha
  • Entrez Gene IDs:
    3559 (Human); 16184 (Mouse); 25704 (Rat); 403870 (Canine)
  • Alternate Names:
    CD25 antigen; CD25; IDDM10; IL-2 R alpha; IL-2 receptor subunit alpha; IL2R alpha; IL-2R subunit alpha; IL2R; IL2RA; IL-2Ra; IL-2-RA; IL2-RA; interleukin 2 receptor, alpha; interleukin-2 receptor subunit alpha; p55; TAC antigen; TCGFR
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