Rabbit anti-Mouse CD25 Antibody | anti-CD25 antibody

Anti-CD25 [7D4], Rabbit IgG

Cat. Num. AAA72279

• Gene Names: Il2ra; CD25; Il2r; Ly-43
• Reactivity: Mouse
• Applications: Flow Cytometry, Functional Assay
• Purity: Protein A affinity purified
• Synonyms: CD25; N/A; Anti-CD25 [7D4], Rabbit IgG; Anti-CD25 [7D4]; IL-2-RA; Interleukin-2 receptor subunit alpha; IL-2 receptor subunit alpha; IL-2R subunit alpha; IL2-RA; p55; anti-CD25 antibody

• Host: Rabbit
• Reactivity: Mouse
• Isotype: Rabbit IgG, Kappa
• Clone Number: 7D4
• Specificity: 7D4 recognizes mouse CD25 at an epitope comprising amino acid 184 to 194 (REHHRFLASEE). This epitope is distinct from the IL-2 binding site and does not block binding of IL-2 to CD25. Murine CD25 is a 55 kDa interleukin-2 receptor alpha chain (IL-2R alpha). CD25 is expressed by early progenitors of the T and B lineage as well as by activated mature T and B lymphocytes. By itself, CD25 binds IL-2 only with low affinity. However, CD25 associates with CD122 (IL-2 receptor beta chain) and CD132 (common gamma chain) to form the high affinity IL-2 receptor.
• Purity/Purification: Protein A affinity purified
• Form/Format: PBS with 0.02% Proclin 300.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-CD25 antibody: FCM/FACS (Flow Cytometry)
• Immunogen: The original rat IgM version of this antibody was raised by immunizing Lewis rats with HT2 cells in complete Freund's adjuvant via a intraperitoneal injection and subsequent generation of hybridomas.
• Chimeric Use Statement: This chimeric rabbit antibody was made using the variable domain sequences of the original Rat IgM format, for improved compatibility with existing reagents, assays and techniques.
• Preparation and Storage: Store at 4 degree C for up to 3 months. For longer storage, aliquot and store at -20 degree C.

IF (Immunofluorescence)

(Immunofluorescence staining of mouse splenocytes with anti-CD25 (AAA72279) 7D4 Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes stained with the chimeric rabbit IgG version of 7D4 (AAA72279-23.0) (1:1000 dilution) for 1h followed by Alexa Fluor 488 secondary antibody (1:1500 dilution), showing nuclear staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom AAA72279-23.0, DAPI, merged channels and an isotype control. The isotype control was anti-fluorescein antibody followed by staining with Alexa Fluor 488 secondary antibody.)

WB (Western Blot)

(Western Blot using anti-CD25 antibody 7D4 (AAA72279). Mouse thymus (0.5ug/ml) tissue lysate (35ug protein in RIPA buffer) was resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of 7D4 (AAA72279-23.0) before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.)

Related Product Information for anti-CD25 antibody

7D4 is a non-IL-2 blocking anti-CD25 antibody that has been widely used to identify CD25-positive cells via flow cytometry (Kohyama et al., 2012; pmid: 22399592), for instance, to analyse Lymph node cells from BALB/c mice that were untreated or treated with 0.25 mg PC61 (Onizuka et al., 1999; pmid: 10397255) or to characterize brain-infiltrating immune cells (Maes et al., 2013). Although 7D4 does not inhibit IL-2 binding itself, it shows synergistic qualities with a cancer vaccine; namely, 7D4 with a cancer vaccine boosted the vaccine-induced anti-tumour response. Furthermore, this antibody was shown to be able to induce inactivation of regulatory T cells (Treg); the effect was particularly significant when this clone was used synergistically with another anti-CD25 clone - PC61 (Kohm et al., 2006; pmid: 16517695; McNeill et al., 2007; pmid: 17212768). 7D4, when turned into a mouse IgG2a format, exhibits depleting qualities against Treg cells and can be used to remove Treg lymphocytes from the tumoral environment without blocking CD25 (IL-2 pathway) on T effector cells (Solomon et al., 2020). As a result, it can be utilized as a potential immunotherapy agent achieving Treg depletion without T effector cells' debilitation (Solomon et al., 2020). Finally, it was demonstrated that such anti-CD25 non-IL-2 blocking antibodies are successful agents in promoting effector activation and leading to strong antitumor activity (Solomon et al., 2020).

Product Categories/Family for anti-CD25 antibody

immunology; Tcells; interleukin

References

Malek et al. 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 Sep;80(18):5694-8. doi: 10.1073/pnas.80.18.5694. PMID:6412230

NCBI and Uniprot Product Information

• NCBI GI #: 124318
• NCBI GeneID: 16184
• UniProt Accession #: P01590; Q61731
• Molecular Weight: 30,723 Da
• NCBI Official Full Name: Interleukin-2 receptor subunit alpha
• NCBI Official Synonym Full Names: interleukin 2 receptor, alpha chain
• NCBI Official Symbol: Il2ra
• NCBI Official Synonym Symbols: CD25; Il2r; Ly-43
• NCBI Protein Information: interleukin-2 receptor subunit alpha; IL2-RA; IL-2-RA; p55 chain; IL-2R alpha chain; IL-2R subunit alpha; IL-2 receptor subunit alpha
• UniProt Protein Name: Interleukin-2 receptor subunit alpha
• UniProt Gene Name: Il2ra
• UniProt Synonym Gene Names: Il2r; IL-2 receptor subunit alpha; IL-2-RA; IL-2R subunit alpha; IL2-RA
• UniProt Entry Name: IL2RA_MOUSE