FITC Rat Anti-Mouse CD25
Clone 7D4 (RUO)
- Brand BD Pharmingen™
- Alternative Name IL-2 Receptor α chain, p55
- Concentration 0.5 mg/ml
- Isotype Rat LEW, also known as Lewis IgM, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen IL-2-dependent BALB/c mouse helper T-cell clone HT-2
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.
FITC, fluorescein isothiocyanate, is a fluorochrome with a molecular weight of 389 Da. FITC is sensitive to pH changes and photobleaching. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. FITC is relatively dim and should be reserved for highly expressed markers whenever possible.
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Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with FITC under optimum conditions, and unreacted FITC was removed.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
For detection of low-density CD25 expression, we recommend the use of the PE conjugate of PC61 antibody (Cat. No. 553866) or the biotin conjugate of the 7D4 antibody (Cat. No. 553069/553070) with a "bright" second-step reagent, such as Streptavidin-PE (Cat. No. 554061).