Alexa Fluor® 647 Rat Anti-Mouse CD25
Clone 7D4 (RUO)
- Brand BD Pharmingen™
- Alternative Name IL-2 Receptor α chain, p55
- Concentration 0.2 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 protein stabilizer and ≤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.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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 fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.