The 7D4 antibody reacts with CD25, the low affinity IL-2 Receptor (IL-2Rα, p55) expressed on activated T and B lymphocytes from all mouse strains tested. IL-2Rα by itself is not a signaling receptor. However, it can combine with IL-2 Receptor β (CD122) and γ (CD132) chains to form high-affinity, signaling receptor complexes for IL-2. Resting T and B lymphocytes and resting and activated NK cells do not express IL-2Rα. CD25 is transiently expressed at a low level during normal B-cell development in the bone marrow on the CD45R/B220low TdT- sIg- Pre-B/Pre-B-II and CD45R/B220low TdT- sIgM+ sIgD- immature B stages, but not on the CD45R/B220low TdT+ sIg- Pro-B/Pre-B-I stage nor on CD45R/B220high TdTsIgM+ sIgD+ mature B cells. It is expressed at a higher level during a very early stage of T-cell development in fetal and adult thymus. Peripheral CD25+ CD4+ T lymphocytes called regulatory T (Treg) cells are involved in the maintenance of self-tolerance. It has also been reported that dendritic cells express CD25, recognized by mAb 7D4 (Cat. no. 553068). The 3C7 antibody recognizes an epitope of CD25 which is distinct from those recognized by mAbs 7D4 and PC61 (Cat. no. 553866), and it blocks binding of IL-2 to CD25. This antibody is routinely tested by flow cytometric analysis. Other applications were tested during antibody development only or reported in the literature. Two-color analysis of the expression of CD25 in bone marrow. BALB/c bone marrow leukocytes were simultaneously stained with biotin-conjugated 7D4 (right panel) and FITC-conjugated RA3-6B2 (anti-mouse CD45R/B220, Cat. No. 553087/553088, both panels) monoclonal antibodies, followed by Streptavidin-PE (Cat. No. 554061, both panels). Flow cytometry was performed on a BD FACScan™
BD Pharmingen™ Biotin Rat Anti-Mouse CD25 - Biotin - Clone 7D4 - Isotype Rat IgM, κ - Reactivity Ms - 0.1 mg
Purification
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
IL2RA
Reactivity: Human
WB, IHC, ELISA, IF
Host: Rabbit
Polyclonal
unconjugated
Application Notes
Flow Cytometry System.
Restrictions
For Research Use only
Concentration
0.5 mg/mL
Buffer
Aqueous buffered solution containing protein stabilizer and ≤0.09 % sodium azide.
Preservative
Sodium azide
Precaution of Use
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
Storage
4 °C
Storage Comment
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Read, Malmström, Powrie: "Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25(+)CD4(+) regulatory cells that control intestinal inflammation." in: The Journal of experimental medicine, Vol. 192, Issue 2, pp. 295-302, (2000) (PubMed).
Takahashi, Tagami, Yamazaki, Uede, Shimizu, Sakaguchi, Mak, Sakaguchi: "Immunologic self-tolerance maintained by CD25(+)CD4(+) regulatory T cells constitutively expressing cytotoxic T lymphocyte-associated antigen 4." in: The Journal of experimental medicine, Vol. 192, Issue 2, pp. 303-10, (2000) (PubMed).
Chen, Ma, Young, Alt: "IL-2 receptor alpha chain expression during early B lymphocyte differentiation." in: International immunology, Vol. 6, Issue 8, pp. 1265-8, (1995) (PubMed).
Rolink, Grawunder, Winkler, Karasuyama, Melchers: "IL-2 receptor alpha chain (CD25, TAC) expression defines a crucial stage in pre-B cell development." in: International immunology, Vol. 6, Issue 8, pp. 1257-64, (1995) (PubMed).
Godfrey, Kennedy, Mombaerts, Tonegawa, Zlotnik: "Onset of TCR-beta gene rearrangement and role of TCR-beta expression during CD3-CD4-CD8- thymocyte differentiation." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 152, Issue 10, pp. 4783-92, (1994) (PubMed).
Taniguchi, Minami: "The IL-2/IL-2 receptor system: a current overview." in: Cell, Vol. 73, Issue 1, pp. 5-8, (1993) (PubMed).
Lorenzo, Jaulin, Vita, Froussard, Ferrara, Jankovic, Theze: "Structure-function study of the p55 subunit of murine IL-2 receptor by epitope mapping." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 147, Issue 9, pp. 2970-7, (1991) (PubMed).
Garni-Wagner, Witte, Tutt, Kuziel, Tucker, Bennett, Kumar: "Natural killer cells in the thymus. Studies in mice with severe combined immune deficiency." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 144, Issue 3, pp. 796-803, (1990) (PubMed).
Pollard, Lipscomb: "Characterization of murine lung dendritic cells: similarities to Langerhans cells and thymic dendritic cells." in: The Journal of experimental medicine, Vol. 172, Issue 1, pp. 159-67, (1990) (PubMed).
Crowley, Inaba, Witmer-Pack, Steinman: "The cell surface of mouse dendritic cells: FACS analyses of dendritic cells from different tissues including thymus." in: Cellular immunology, Vol. 118, Issue 1, pp. 108-25, (1989) (PubMed).
Habu, Okumura, Diamantstein, Shevach: "Expression of interleukin 2 receptor on murine fetal thymocytes." in: European journal of immunology, Vol. 15, Issue 5, pp. 456-60, (1985) (PubMed).
Malek, Schmidt, Shevach: "The murine IL 2 receptor. III. Cellular requirements for the induction of IL 2 receptor expression on T cell subpopulations." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 134, Issue 4, pp. 2405-13, (1985) (PubMed).
Ortega, Robb, Shevach, Malek: "The murine IL 2 receptor. I. Monoclonal antibodies that define distinct functional epitopes on activated T cells and react with activated B cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 133, Issue 4, pp. 1970-5, (1984) (PubMed).
Malek, Robb, Shevach: "Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 80, Issue 18, pp. 5694-8, (1983) (PubMed).