Anti-Mouse CD25 Antibody, Clone 3C7

Rat monoclonal IgG2b antibody against mouse CD25

产品号 #60010_C

Rat monoclonal IgG2b antibody against mouse CD25

概述

The 3C7 antibody reacts with murine CD25 (interleukin‐2 receptor α chain; IL‐2Rα), an ~55 kDa type 1 transmembrane glycoprotein expressed on the surface of T and B cell progenitors and activated mature B and T cells. CD25 has been used as a marker to identify CD4+/FoxP3+ regulatory T cells in mice. Mutations in CD25 result in severe immunodeficiency, underscoring the importance of the roles the protein plays in the differentiation, activation and proliferation of lymphocytes, and in the maintenance of self-tolerance. CD25 per se has low affinity for its IL-2 ligand but forms dimers with CD122 (IL-2Rβ) and CD132 (IL-2Rγ) that together associate to form the IL-2R receptor, which binds IL-2 with high affinity. CD25 acts to increase the specificity and affinity of IL-2 binding and is necessary for receptor clustering and signal transduction by the complex. Binding of the 3C7 antibody inhibits binding of IL‐2 to free CD25 and the IL-2R receptor. The epitope is distinct from those recognized by the anti-CD25 antibody clones 7D4 and PC61.

This antibody clone has been verified for purity assessments of cells isolated with EasySep™ kits, including EasySep™ Mouse T Cell Enrichment Kit (Catalog #19751).

Subtype
Primary Antibodies
 
Target Antigen
CD25
 
Alternative Names
IL-2Ralpha, Ly-43, p55, Tac
 
Reactive Species
Mouse
 
Conjugation
FITC, PE, Unconjugated
 
Host Species
Rat
 
Cell Type
T Cells, T Cells, Regulatory
 
Species
Mouse
 
Application
Cell Isolation, CyTOF, Flow Cytometry, Functional Assay, Immunohistochemistry
 
Area of Interest
Immunology
 
Clone
3C7
 
Gene ID
16184
 
Isotype
IgG2b, kappa
 

Data Figures

Data for Unconjugated

Figure 1. Data for Unconjugated

(A) Flow cytometry analysis of C57BL/6 mouse splenocytes (gated on CD3e+ cells) labeled with Anti-Mouse CD25 Antibody, Clone 3C7, followed by a mouse anti-rat IgG2b antibody, FITC and Anti-Mouse CD4, Clone RM4-5, PE (Catalog #60017PE). (B) Flow cytometry analysis of C57BL/6 mouse splenocytes (gated on CD3e+ cells) labeled with Rat IgG2b, kappa Isotype Control Antibody, Clone RTK4530 (Catalog #60077), followed by a mouse anti-rat IgG2b antibody, FITC and Anti-Mouse CD4, Clone RM4-5, PE.

Data for FITC-Conjugated

Figure 2. Data for FITC-Conjugated

(A) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Anti-Mouse CD25 Antibody, Clone 3C7, FITC and anti-mouse CD3 APC.
(B) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with a rat IgG2b, kappa FITC isotype control antibody and anti-mouse CD3 APC.
(C) Flow cytometry analysis of mouse T cells activated with antibodies against CD3 and CD28. C57BL/6 mouse splenocytes were processed with the EasySep™ Mouse T Cell Enrichment Kit and the enriched T cells were cultured in the absence (Unstimulated) or presence (Stimulated) of Anti-Mouse CD28 and plate-bound Anti-Mouse CD3e for 48 hours, then labeled with Anti-Mouse CD25 Antibody, Clone 3C7, FITC and analyzed for CD25 expression. Upregulation of CD25 following stimulation with CD3 and CD28 is apparent (Stimulated). Labeling of C57BL/6 mouse splenocytes with a rat IgG2b, kappa Alexa Fluor® 488 isotype control antibody is shown (open histogram).

Data for PE-Conjugated

Figure 3. Data for PE-Conjugated

(A) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Anti-Mouse CD25 Antibody, Clone 3C7, PE and anti-mouse CD3 APC.
(B) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with a rat IgG2b, kappa PE isotype control antibody and anti-mouse CD3 APC.
(C) Flow cytometry analysis of mouse T cells activated with antibodies against CD3 and CD28. C57BL/6 mouse splenocytes were processed with the EasySep™ Mouse T Cell Enrichment Kit (Catalog #19751) and the enriched T cells were cultured in the absence (Unstimulated) or presence (Stimulated) of Anti-Mouse CD28 and plate-bound Anti-Mouse CD3e for 48 hours, then labeled with Anti-Mouse CD25 Antibody, Clone 3C7, PE and analyzed for CD25 expression. Upregulation of CD25 following stimulation with CD3 and CD28 is apparent (Stimulated). Labeling of C57BL/6 mouse splenocytes with a rat IgG2b, kappa Alexa Fluor® 488 isotype control antibody is shown (open histogram).

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
60010FI.1
Lot #
All
Language
English
Catalog #
60010PE
Lot #
All
Language
English
Catalog #
60010
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
60010FI.1
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
60010PE
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
60010
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Resources and Publications

Educational Materials (2)

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