Skip to main content Skip to navigation
PE Mouse Anti-Mouse Foxp3
PE Mouse Anti-Mouse Foxp3

Multicolor flow cytometric analysis of Foxp3 expression in mouse splenocytes. C57BL/6 mouse spleen cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed, fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD4 antibody (Cat. No. 562891), APC Rat Anti-Mouse CD25 antibody (Cat. No. 557192) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Top Plots) or PE Mouse Anti-Foxp3 antibody (Cat. No. 566881; Bottom plots) at 0.125 µg/test.

     Left Plots: Two-color flow cytometric dot plots showing the correlated expression of CD4 versus Foxp3 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact cells.

     Right Plots: Two-color flow cytometric dot plots showing the correlated expression of Foxp3 (or Ig Isotype control staining) versus CD25 were derived from CD4 positive-gated events with the light-scatter characteristics of intact cells.

     Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Multicolor flow cytometric analysis of Foxp3 expression in mouse splenocytes. C57BL/6 mouse spleen cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed, fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD4 antibody (Cat. No. 562891), APC Rat Anti-Mouse CD25 antibody (Cat. No. 557192) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Top Plots) or PE Mouse Anti-Foxp3 antibody (Cat. No. 566881; Bottom plots) at 0.125 µg/test.

     Left Plots: Two-color flow cytometric dot plots showing the correlated expression of CD4 versus Foxp3 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact cells.

     Right Plots: Two-color flow cytometric dot plots showing the correlated expression of Foxp3 (or Ig Isotype control staining) versus CD25 were derived from CD4 positive-gated events with the light-scatter characteristics of intact cells.

     Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
Down Arrow Up Arrow


BD Pharmingen™
Foxp3; JM2; scurfin; scurfy; sf
Mouse (QC Testing)
Mouse BALB/c IgG1, κ
Mouse Foxp3 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2869932
Aqueous buffered solution containing ≤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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566881 Rev. 1
Antibody Details
Down Arrow Up Arrow
3G3

The 3G3 monoclonal antibody specifically binds to Forkhead box protein P3 (Foxp3) which is also known as Scurfin and JM2. Foxp3 is a 50-55 kDa protein that is encoded by Foxp3 (forkhead box P3) which belongs to the forkhead/winged helix family of transcriptional regulators. FoxP3 contains a single C2H2-type zinc-finger motif, a leucine-zipper region, and a C-terminal forkhead DNA-binding domain. Foxp3 is expressed by natural (nTreg) and induced/adaptive (iTreg) T regulatory cells. Foxp3 is a key transcription factor for Treg cell development and regulatory function. Treg cells play crucial roles in maintaining immune homeostasis by enforcing immunological tolerance to self-antigens and by suppressing excessive responses made by other immune cells to foreign antigens. Ectopic expression of Foxp3 in conventional T cells is sufficient to induce suppressive activity, repress the production of cytokines such as IL2 and IFN-γ, and upregulate Treg cell-associated molecules such as CTLA4/CD152 and GITR/TNFRSF18. A mutation in Foxp3 causes a lack of functional Tregs in Scurfy (sf) mice which develop a systemic disease state with autoimmune manifestations. The 3G3 antibody reportedly binds to the N-terminus of Foxp3.

        

566881 Rev. 1
Format Details
Down Arrow Up Arrow
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566881 Rev.1
Citations & References
Down Arrow Up Arrow

Development References (5)

  1. Brunkow ME, Jeffery EW, Hjerrild KA, et al. Disruption of a new forkhead/winged-helix protein, scurfin, results in the fatal lymphoproliferative disorder of the scurfy mouse. Nat Genet. 2001; 27(1):68-73. (Biology). View Reference
  2. Gavin MA, Torgerson TR, Houston E, et al. Single-cell analysis of normal and FOXP3-mutant human T cells: FOXP3 expression without regulatory T cell development. Proc Natl Acad Sci U S A. 2006; 103(17):6659-6664. (Immunogen: Flow cytometry). View Reference
  3. Hadaschik EN, Wei X, Leiss H, et al. Regulatory T cell-deficient scurfy mice develop systemic autoimmune features resembling lupus-like disease. Arthritis Res Ther. 2015; 17(35):1-12. (Biology). View Reference
  4. Law JP, Hirschkorn DF, Owen RE, Biswas HH, Norris PJ, Lanteri MC. The importance of Foxp3 antibody and fixation/permeabilization buffer combinations in identifying CD4+CD25+Foxp3+ regulatory T cells. Cytometry A. 2009; 75(12):1040-1050. (Clone-specific: Flow cytometry). View Reference
  5. Mailer RKW. Alternative Splicing of FOXP3-Virtue and Vice. Front Immunol. 2018; 9(530):1-11. (Clone-specific). View Reference
View All (5) View Less
566881 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.