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Biotin Rat Anti-Mouse CD25
Biotin Rat Anti-Mouse CD25

Immunohistochemical staining of CD25 positive cells. Frozen sections of mouse thymus were reacted with Biotin Rat Anti-Mouse CD25 antibody (Cat. No. 550529). Immature thymocytes expressing the IL-2 receptor can be identified by the brown labeling of their cell membranes. Amplification 20x.

Immunohistochemical staining of CD25 positive cells. Frozen sections of mouse thymus were reacted with Biotin Rat Anti-Mouse CD25 antibody (Cat. No. 550529). Immature thymocytes expressing the IL-2 receptor can be identified by the brown labeling of their cell membranes. Amplification 20x.

Product Details
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BD Pharmingen™
IL-2 Receptor α chain, p55
Mouse (QC Testing)
Rat LEW, also known as Lewis IgM, κ
IL-2-dependent BALB/c mouse helper T-cell clone HT-2
Flow cytometry (Routinely Tested), Immunohistochemistry-frozen (Tested During Development), Immunohistochemistry-paraffin (Not Recommended)
125 µg/ml
AB_2125455
Aqueous buffered solution containing BSA, protein stabilizer, goat serum, and ≤ 0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.

Recommended Assay Procedures

Immunohistochemistry: The 7D4 antibody specific for mouse CD25 is recommended to test for immunohistochemical staining of acetone-fixed frozen sections. Tissues tested were mouse spleen and thymus. The antibody stains activated T and B lymphocytes and immature thymocytes. The isotype control recommended for use with this antibody is Biotin Rat IgM, κ Isotype Control (Cat. No. 550343). For optimal indirect immunohistochemical staining, the biotin format of the 7D4 antibody should be titrated (1:10 to 1:50 dilution) and visualized by Streptavidin HRP (Cat. No. 550946) together with the DAB Substrate Kit (Cat. No. 550880). A detailed protocol of the immunohistochemical procedure can  be found on our website at http://www.bdbiosciences.com/us/s/resources. The clone 7D4 is not recommended for form alin-fixed paraffin embedded sections.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  5. Use of these products to measure activation antigens expressed on mononuclear cell subsets for the purpose of monitoring immunoregulatory status can fall under one or more claims of the following patents: US Patent Nos. 5,445,939, 5,656,446, 5,843,689; European Patent No. 319,543; Canadian Patent No. 1,296,622; Australian Patent No. 615,880; and Japanese Patent No. 2,769,156.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
550529 Rev. 3
Antibody Details
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7D4

The 7D4 monoclonal antibody specifically binds to 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 as well as 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, which is recognized by mAb 7D4. The 3C7 antibody recognizes an epitope of CD25 which is distinct from those recognized by mAbs 7D4 and PC61, and it blocks binding of IL-2 to CD25.

550529 Rev. 3
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
550529 Rev.3
Citations & References
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Development References (10)

  1. Chen J, Ma A, Young F, Alt FW. IL-2 receptor alpha chain expression during early B lymphocyte differentiation. Int Immunol. 1994; 6(8):1265-1268. (Biology). View Reference
  2. Crowley M, Inaba K, Witmer-Pack M, Steinman RM. The cell surface of mouse dendritic cells: FACS analyses of dendritic cells from different tissues including thymus. Cell Immunol. 1989; 118(1):108-125. (Clone-specific: Cytotoxicity). View Reference
  3. Garni-Wagner BA, Witte PL, Tutt MM, et al. Natural killer cells in the thymus. Studies in mice with severe combined immune deficiency. J Immunol. 1990; 144(3):796-803. (Biology). View Reference
  4. Godfrey DI, Kennedy J, Mombaerts P, Tonegawa S, Zlotnik A. Onset of TCR-β gene rearrangement and role of TCR-β expression during CD3-CD4-CD8- thymocyte differentiation. J Immunol. 1994; 152(10):4783-4792. (Biology). View Reference
  5. Habu S, Okumura K, Diamantstein T, Shevach EM. Expression of interleukin 2 receptor on murine fetal thymocytes. Eur J Immunol. 1985; 15(5):456-460. (Clone-specific: Immunohistochemistry). View Reference
  6. Malek TR, Robb RJ, Shevach EM. Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex. Proc Natl Acad Sci U S A. 1983; 80(18):5694-5698. (Immunogen: Flow cytometry, Immunoprecipitation, Inhibition). View Reference
  7. Malek TR, Schmidt JA, Shevach EM. The murine IL 2 receptor. III. Cellular requirements for the induction of IL 2 receptor expression on T cell subpopulations. J Immunol. 1985; 134(4):2405-2413. (Biology). View Reference
  8. Ortega G, Robb RJ, Shevach EM, Malek TR. The murine IL 2 receptor. I. Monoclonal antibodies that define distinct functional epitopes on activated T cells and react with activated B cells. J Immunol. 1984; 133(4):1970-1975. (Clone-specific: Immunoprecipitation, Inhibition). View Reference
  9. Pollard AM, Lipscomb MF. Characterization of murine lung dendritic cells: similarities to Langerhans cells and thymic dendritic cells. J Exp Med. 1990; 172(1):159-167. (Clone-specific: Cytotoxicity). View Reference
  10. Taniguchi T, Minami Y. The IL-2/IL-2 receptor system: a current overview. Cell. 1993; 73(1):5-8. (Biology). View Reference
View All (10) View Less
550529 Rev. 3

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