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PerCP-Cy™5.5 Rat Anti-Mouse Foxp3
PerCP-Cy™5.5 Rat Anti-Mouse Foxp3
Multicolor flow cytometric analysis of Foxp3 expression in mouse splenocytes. Mouse splenic leucocytes were fixed and permeabilized with the Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553047/553046/561835) and either PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; Left Plot) or PerCP-Cy5.5 Rat Anti-Mouse Foxp3 (Cat. No. 563902; Right Plot) antibodies at 0.5 µg/test. The bivariate pseudocolor density plot shows the correlated expression patterns of Foxp3 (or Ig Isotype control staining) versus CD4 for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data was performed using a BD™ LSR II Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
PerCP-Cy™5.5 Rat Anti-Mouse Foxp3
Two-color flow cytometric analysis of Foxp3 expression in mouse splenocytes.  Mouse splenic leucocytes were fixed and permeabilized using appropriately-diluted solutions from the Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553047/553046/561835) and PerCP-Cy™5.5 Rat Anti-Mouse Foxp3 (Cat. No. 563902) antibodies. The two-color flow cytometric dot plot shows the correlated expression patterns of Foxp3 versus CD4 for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of Foxp3 expression in mouse splenocytes. Mouse splenic leucocytes were fixed and permeabilized with the Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553047/553046/561835) and either PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; Left Plot) or PerCP-Cy5.5 Rat Anti-Mouse Foxp3 (Cat. No. 563902; Right Plot) antibodies at 0.5 µg/test. The bivariate pseudocolor density plot shows the correlated expression patterns of Foxp3 (or Ig Isotype control staining) versus CD4 for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data was performed using a BD™ LSR II Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of Foxp3 expression in mouse splenocytes.  Mouse splenic leucocytes were fixed and permeabilized using appropriately-diluted solutions from the Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553047/553046/561835) and PerCP-Cy™5.5 Rat Anti-Mouse Foxp3 (Cat. No. 563902) antibodies. The two-color flow cytometric dot plot shows the correlated expression patterns of Foxp3 versus CD4 for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
Forkhead box P3; IPEX; Forkhead box protein P3; JM2; Scurfin; Scurfy; Sf
Mouse (QC Testing)
Rat IgG2a, κ
Foxp3 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2630318
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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. An isotype control should be used at the same concentration as the antibody of interest.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  5. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  6. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
563902 Rev. 4
Antibody Details
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R16-715

The R16-715 monoclonal antibody specifically binds to mouse Foxp3. Foxp3 is a 50-55 kDa protein also known as Forkhead box P3, JM2, IPEX, Scurfin, and  Sf. It is a member of the forkhead or winged helix family of transcription factors and is specifically expressed by T regulatory (Treg) cells. Foxp3 is a key regulatory protein for Treg cell development and function. 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 CD25, CTLA4 and GITR. It has been found that the mutation of Foxp3 is responsible for "scurfy" mice. When overexpressed, Foxp3 leads to poor T cell proliferation and activation.

563902 Rev. 4
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
563902 Rev.4
Citations & References
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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. Hori S, Nomura T, Sakaguchi S. Control of regulatory T cell development by the transcription factor Foxp3. Science. 2003; 299(5609):1057-1061. (Biology). View Reference
  3. Jinushi M, Sato M, Kanamoto A, et al. Milk fat globule epidermal growth factor-8 blockade triggers tumor destruction through coordinated cell-autonomous and immune-mediated mechanisms. J Exp Med. 2009; 206(6):1317-1326. (Biology). View Reference
  4. Vasconcellos R, Carter NA, Rosser EC, Mauri C. IL-12p35 subunit contributes to autoimmunity by limiting IL-27-driven regulatory responses. J Immunol. 2011; 187(6):3402-3412. (Biology). View Reference
  5. Zheng Y, Rudensky AY. Foxp3 in control of the regulatory T cell lineage. Nat Immunol. 2007; 8:457-462. (Biology). View Reference
View All (5) View Less
563902 Rev. 4

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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.