Anti-CD25 [2E4]

Name: Anti-CD25 [2E4]
SKU: Ab02358
Availability: InStock

Description:

Recombinant monoclonal antibody to CD25. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 2E4.

Clone Name:

2E4

Published Application(s):

Depletion, FC

Published Species Reactivity:

Mouse

Products based on this clone:

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Ab02358-1.1 Anti-CD25 [2E4]
Mouse
Mouse IgG1
Purified
Ships in 4-5 weeks

Ab02358-2.0 Anti-CD25 [2E4]
Mouse
Mouse IgG2a
Purified
In Stock

Ab02358-2.3 Anti-CD25 [2E4]
Mouse
Mouse IgG2a
Fc Silent™
Purified
Ships in 4-5 weeks

Ab02358-23.0 Anti-CD25 [2E4]
Mouse
Rabbit IgG
Purified
In Stock

Ab02358-10.0 Anti-CD25 [2E4]
Mouse
Human IgG1
Purified
In Stock

Ab02358-15.0 Anti-CD25 [2E4]
Mouse
Human IgM
Purified
In Stock

UniProt Accession Number of Target Protein: P01590

Alternative Name(s) of Target: IL-2-RA; Interleukin-2 receptor subunit alpha; IL-2 receptor subunit alpha; IL-2R subunit alpha; IL2-RA; p55
Immunogen: The original version of this antibody was raised to recognize mouse CD25.

Specificity: 2E4 recognizes mouse CD25. This epitope is distinct from the IL-2 binding site and does not block binding of IL-2 to CD25. This antibody binds the same or overlapping epitope as 7D4 (Ab02216) but different than Ab01107. 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.

Application Notes: 2E4 is a non-IL-2 blocking anti-CD25 antibody. The variable fragment (Fv) of this antibody conjugated to a 38-kDa portion of Pseudomonas exotoxin A exhibited exceptional ability to induce tumor rejection and lasting antitumor immunitiy in mice making it a promising canditate in the immunotherapy research and developemnt (Onda et al., 2019; pmid: 30760587). This 2E4 Fv-based CD25-specific recombinant immunotoxin (RIT) was injected into three different mouse tumor models and brought about complete regressions of the majority of them leadinig, in many cases, to immunity against tumors' reinjections and cross-tumor type immunity (Onda et al., 2019; pmid: 30760587). Finally, it was demonstrated that this immunotoxin (RIT) caused significant depletion of regulatory T cells (Tregs) in the tumor environment but, importantly, not in the spleen; at the same time, the number of effector, cytotoxic CD8 cells in the tumor increased what would suggest the possible antitumour mechanism of this immunotoxin (Onda et al., 2019; pmid: 30760587). Additionally, 2E4 can be used in flow cytometric analyses; for instance, it has been used to perform phenotyping of lymphocytes as a part of a wider panel of antibodies (Alexander et al., 2008; pmid: 18824594).

Antibody first published in: Ortega et al. 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 Oct;133(4):1970-5. PMID:6206144
Note on publication: Describes the characterization of the antibody 2E4.

Flow cytometry analysis of mouse splenocytes using Ab02358-23.0. Click on image for more details.

Flow cytometry using the Anti-CD25 antibody 2E4 (Ab02358). Paraformaldehyde fixed mouse splenocytes permeabilized with 0.5% Triton were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of 2E4 (Ab02358-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

Western blot on mouse lymph node tissue lysate using Ab02358-23.0. Click on image for more details.

Western Blot using Anti-CD25 antibody 2E4 (Ab02358). Mouse lymph node tissue lysates (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of 2E4 (Ab02358-23.0) at 0.5 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.

IF staining of mouse splenocytes using Ab02358-23.0. Click on image for more details.

Immunofluorescence staining of mouse splenocytes with anti-CD25 (Ab02358) 2E4 Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes on Shi-fix™ cover-slips stained with the chimeric rabbit IgG version of 2E4 (Ab02358-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab02358-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.