MABF2101

Anti-CD25 Antibody, clone 7D4

• UNSPSC Code: 12352203

Properties

• biological source: rat
• Quality Level: 200
• conjugate: unconjugated
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 7D4, monoclonal
• mol wt: calculated mol wt 30.72 kDa
• purified by: ion-exchange chromatography
• species reactivity: mouse
• packaging: antibody small pack of 100 μg
• technique(s): flow cytometry: suitable; immunocytochemistry: suitable; immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable; immunoprecipitation (IP): suitable; inhibition assay: suitable; radioimmunoassay: suitable
• isotype: IgM
• epitope sequence: Extracellular domain
• Protein ID accession no.: NP_032393
• UniProt accession no.: P01590
• shipped in: dry ice
• target post-translational modification: unmodified

Description

General description

Interleukin-2 receptor subunit alpha (UniProt: P01590; also known as IL-2 receptor subunit alpha, IL-2-RA, IL-2R subunit alpha, IL2-RA, p55, CD25) is encoded by the Il2ra (also known as Il2r) gene (Gene ID: 16184) in murine species. CD25 is a single-pass type I membrane glycoprotein that is synthesized with a signal peptide (aa 1-21), which is subsequently cleaved off to produce the mature form that contains an extracellular domain (aa 22-236), a transmembrane domain (aa 237-257), and a short cytoplasmic domain (aa 258-268). CD25 is mainly expressed on activated T -cells where it serves as a receptor for Interleukin-2 and is involved in the regulation of immune tolerance by controlling regulatory T cells (TREGs) activity that suppresses the activation and expansion of autoreactive T-cells. The IL-2 receptor (IL-2R) is composed of three different subunit complexes, the a- (CD25), β- (CD122) and g-chains (CD132). By itself, CD25 is not sufficient to trigger the IL-2 signaling pathway because it is a low-affinity form of the IL-2 receptor (KD = 10 8 M) that is non-functional with respect to IL-2 internalization and signaling due to its short cytoplasmic tail. It is mainly involved in IL-2 binding thereby stabilizing a secondary binding site for the β-chain and recruiting g-chain to the cell surface. Clone 7D4 is shown to down-modulate the expression of CD25 on CD4+ T cells but does not affect the level of Foxp3+ T cells. It is also a potent inhibitor of IL 2- driven proliferation of HT2 or CTL-L cells. (Ref.: Nihei, J., et al. (2014). Eur. J. Microbiol. Immunol. 4(2); 128-137; Ortega, G., et al. (1984). J. Immunol. 133(4); 1970-1975).

Specificity

Clone 7D4 is a rat monoclonal antibody that detects murine Interleukin-2 receptor subunit alpha (CD25). It targets an epitope within the Sushi-2 domain in the extracellular region.

Immunogen

IL-2 dependent murine T helper HT2 cells.

Application

Quality Control Testing

Evaluated by Immunocytochemistry in RAW264.7 cells.

Immunocytochemistry Analysis (ICC): 1:25 dilution of this antibody detected Interleukin-2 receptor subunit alpha (CD25) in RAW264.7 cells.

Tested Applications

Inhibition: A representative lot inhibited proliferation of the HT2 cells and of IL-2-dependent CTLL cells. (Malek, T.R., et al. (1983). Proc Natl Acad Sci USA.;80(18):5694-8; Ortega, G., et al. (1984). J Immunol.;133(4):1970-5; Pollard, A.M., et al. (1990). J Exp Med.;172(1):159-67; Nihei, J. et al. (2014). Eur J Microbiol Immunol (Bp).;4(2):128-37; Teege, S., et al. (2015). Sci Rep.;5:8959).

Immunohistochemistry Applications: A representative lot detected CD25 in Immunohistochemistry applications (Nihei, J. et al. (2014). Eur J Microbiol Immunol (Bp).;4(2):128-37).

Immunoprecipitation Analysis: A representative lot immunoprecipitated CD25 in Immunoprecipitation applications (Malek, T.R., et al. (1983). Proc Natl Acad Sci USA.;80(18):5694-8; Ortega, G., et al. (1984). J Immunol.;133(4):1970-5).

Flow Cytometry: A representative lot detected CD25 in Flow Cytometry applications (Malek, T.R., et al. (1983). Proc Natl Acad Sci USA.;80(18):5694-8; Ortega, G., et al. (1984). J Immunol.;133(4):1970-5; Malek, T.R., et al. (1985). J Immunol.;134(4):2405--13; Kohm, A.P., et al. (2006). J Immunol.;176(6):3301-5; Nihei, J. et al. (2014). Eur J Microbiol Immunol (Bp).;4(2):128-37; Teege, S., et al. (2015). Sci Rep.;5:8959).

Radioimmunoassay: A representative lot detected CD25 in Radioimmunoassay applications (Ortega, G., et al. (1984). J Immunol.;133(4):1970-5).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-CD25, clone 7D4, Cat. No. MABF2101, is a rat monoclonal antibody that detects CD25 and is tested for use in Flow Cytometry, Immunocytochemistry, Immunohistochemistry, Immunoprecipitation, Inhibition Assay, and Radioimmunoassay.

Physical form

Purified rat monoclonal antibody IgM in PBS without preservatives.

Storage and Stability

Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 2
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable