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BD Horizon™ BV421 Rat Anti-Mouse CD25
Clone 3C7 (RUO)
Flow cytometric analysis of CD25 expression on unstimulated and stimulated mouse splenocytes.
Left and Middle Panels: Freshly prepared mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051/561091) and either BD Horizon™ BV421 Rat IgG2b, κ Isotype Control (Cat. No. 562603; Left Panel) or BD Horizon BV421 Rat Anti-Mouse CD25 antibody (Cat. No. 564370; Middle Panel). Two-color flow cytometric contour plots showing the correlated expression patterns for CD25 (or Ig Isotype Control staining) versus CD4 were generated for gated events with the forward and side light- scatter characteristics of viable lymphocytes.
Right Panel: Mouse splenic leucocytes were stimulated with concanavalin A for 3 days. The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with either BD Horizon BV421 Rat IgG2b, κ Isotype Control (dashed line histogram) or BD Horizon BV421 Rat Anti-Mouse CD25 antibody (solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphoblasts.
Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated mouse splenocytes.
Left and Middle Panels: Freshly prepared mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051/561091) and either BD Horizon™ BV421 Rat IgG2b, κ Isotype Control (Cat. No. 562603; Left Panel) or BD Horizon BV421 Rat Anti-Mouse CD25 antibody (Cat. No. 564370; Middle Panel). Two-color flow cytometric contour plots showing the correlated expression patterns for CD25 (or Ig Isotype Control staining) versus CD4 were generated for gated events with the forward and side light- scatter characteristics of viable lymphocytes.
Right Panel: Mouse splenic leucocytes were stimulated with concanavalin A for 3 days. The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with either BD Horizon BV421 Rat IgG2b, κ Isotype Control (dashed line histogram) or BD Horizon BV421 Rat Anti-Mouse CD25 antibody (solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphoblasts.
Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated mouse splenocytes.
Left and Middle Panels: Freshly prepared mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051/561091) and either BD Horizon™ BV421 Rat IgG2b, κ Isotype Control (Cat. No. 562603; Left Panel) or BD Horizon BV421 Rat Anti-Mouse CD25 antibody (Cat. No. 564370; Middle Panel). Two-color flow cytometric contour plots showing the correlated expression patterns for CD25 (or Ig Isotype Control staining) versus CD4 were generated for gated events with the forward and side light- scatter characteristics of viable lymphocytes.
Right Panel: Mouse splenic leucocytes were stimulated with concanavalin A for 3 days. The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with either BD Horizon BV421 Rat IgG2b, κ Isotype Control (dashed line histogram) or BD Horizon BV421 Rat Anti-Mouse CD25 antibody (solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphoblasts.
Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated mouse splenocytes.
Left and Middle Panels: Freshly prepared mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051/561091) and either BD Horizon™ BV421 Rat IgG2b, κ Isotype Control (Cat. No. 562603; Left Panel) or BD Horizon BV421 Rat Anti-Mouse CD25 antibody (Cat. No. 564370; Middle Panel). Two-color flow cytometric contour plots showing the correlated expression patterns for CD25 (or Ig Isotype Control staining) versus CD4 were generated for gated events with the forward and side light- scatter characteristics of viable lymphocytes.
Right Panel: Mouse splenic leucocytes were stimulated with concanavalin A for 3 days. The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with either BD Horizon BV421 Rat IgG2b, κ Isotype Control (dashed line histogram) or BD Horizon BV421 Rat Anti-Mouse CD25 antibody (solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphoblasts.
Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Horizon™
Interleukin-2 receptor alpha chain; IL-2RA; IL-2Rα; Il2ra; IL-2R p55
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2b, κ
IL-2-dependent BALB/c mouse cell line
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2738772
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
Product Notices
- 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.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
BV421 Rat IgG2b, κ Isotype Control RUO
Size
50 µg
Cat No.
562603
564370 Rev. 1
Antibody Details
3C7
The 3C7 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 γc (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 during the CD45R/B220low TdT- sIg- Pre-B/Pre-B-II and CD45R/B220low TdT- sIgM+ sIgD- immature B stages, but not during 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 (recognized by mAb 7D4, another CD25-specific antibody). The 3C7 antibody recognizes an epitope of CD25 which is distinct from those recognized by the other CD25-specific mAbs, 7D4 and PC61. 3C7 blocks the binding of IL-2 to CD25.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
564370 Rev. 1
Format Details
BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
407 nm
423 nm
564370 Rev.1
Citations & References
View product citations for antibody "564370" on CiteAb
Development References (12)
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Chen J, Ma A, Young F, Alt FW. IL-2 receptor alpha chain expression during early B lymphocyte differentiation. Int Immunol. 1994; 6(8):1265-1268. (Clone-specific: Flow cytometry). View Reference
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Crowley M, Inaba K, Witmer-Pack M, Steinman RM. The cell surface of mouse dendritic cells: FACS analyses of dendritic cells from different tissues including thymus. Cell Immunol. 1989; 118(1):108-125. (Biology). View Reference
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Garni-Wagner BA, Witte PL, Tutt MM, et al. Natural killer cells in the thymus. Studies in mice with severe combined immune deficiency. J Immunol. 1990; 144(3):796-803. (Biology). View Reference
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Habu S, Okumura K, Diamantstein T, Shevach EM. Expression of interleukin 2 receptor on murine fetal thymocytes. Eur J Immunol. 1985; 15(5):456-460. (Biology). View Reference
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Malek TR, Robb RJ, Shevach EM. 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; 80(18):5694-5698. (Biology). View Reference
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Malek TR, Schmidt JA, Shevach EM. The murine IL 2 receptor. III. Cellular requirements for the induction of IL 2 receptor expression on T cell subpopulations. J Immunol. 1985; 134(4):2405-2413. (Biology). View Reference
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Malek TR. The biology of interleukin-2. Annu Rev Immunol. 2008; 26:453-479. (Biology). View Reference
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Moreau JL, Nabholz M, Diamantstein T, Malek T, Shevach E, Theze J. Monoclonal antibodies identify three epitope clusters on the mouse p55 subunit of the interleukin 2 receptor: relationship to the interleukin 2-binding site. Eur J Immunol. 1987; 17(7):929-935. (Clone-specific: Bioassay, Blocking, Functional assay, Inhibition, Neutralization, Radioimmunoassay). View Reference
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Ortega G, Robb RJ, Shevach EM, Malek TR. 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; 133(4):1970-1975. (Immunogen: Blocking, Flow cytometry, Immunoprecipitation, Inhibition, Neutralization, Radioimmunoassay). View Reference
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Pollard AM, Lipscomb MF. Characterization of murine lung dendritic cells: similarities to Langerhans cells and thymic dendritic cells. J Exp Med. 1990; 172(1):159-167. (Biology). View Reference
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Read S, Malmstrom V, Powrie F. Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25(+)CD4(+) regulatory cells that control intestinal inflammation. J Exp Med. 2000; 192(2):295-302. (Biology). View Reference
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Rolink A, Grawunder U, Winkler TH, Karasuyama H, Melchers F. IL-2 receptor alpha chain (CD25, TAC) expression defines a crucial stage in pre-B cell development. Int Immunol. 1994; 6(8):1257-1264. (Biology). View Reference
564370 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.