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PE-CF594 Rat Anti-Mouse Foxp3
PE-CF594 Rat Anti-Mouse Foxp3
Multicolor flow cytometric analysis of Foxp3 expression in BALB/c mouse splenocytes. Splenocytes from BALB/c mice were fixed and permeabilized using working solutions of Mouse Foxp3 Buffers (Mouse Foxp3 Buffer Set, Cat. No. 560409). The cells were then stained with PerCP-Cy™5.5 Rat anti-Mouse CD4 (Cat. No. 550954/561115) and BD Horizon™ PE-CF594 Rat anti-Mouse Foxp3 antibodies (Cat. No. 562466). The flow cytometric dot plot shows the correlated expression of Foxp3 versus CD4 for gated events with the forward and side light-scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.
Multicolor flow cytometric analysis of Foxp3 expression in BALB/c mouse splenocytes. Splenocytes from BALB/c mice were fixed and permeabilized using working solutions of Mouse Foxp3 Buffers (Mouse Foxp3 Buffer Set, Cat. No. 560409). The cells were then stained with PerCP-Cy™5.5 Rat anti-Mouse CD4 (Cat. No. 550954/561115) and BD Horizon™ PE-CF594 Rat anti-Mouse Foxp3 antibodies (Cat. No. 562466). The flow cytometric dot plot shows the correlated expression of Foxp3 versus CD4 for gated events with the forward and side light-scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.
Product Details
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BD Horizon™
Forkhead box P3; Forkhead box protein P3; JM2; Scurfin; Scurfy; Sf
Mouse (QC Testing)
Rat IgG2b, κ
Mouse FoxP3 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_11151905
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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.

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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  8. CF™ is a trademark of Biotium, Inc.
  9. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  10. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  11. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  13. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
562466 Rev. 4
Antibody Details
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MF23

Foxp3 is a 50-55 kDa protein also known as Forkhead box P3, JM2, or IPEX. It is a member of the forkhead or winged helix family of

transcription factors and is specifically expressed by T regulatory (Treg) cells. Foxp3 has been reported to be 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

562466 Rev. 4
Format Details
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, 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 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-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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PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
562466 Rev.4
Citations & References
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Development References (5)

  1. 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
  2. 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. (Clone-specific: Flow cytometry). View Reference
  3. Ono M, Yaguchi H, Ohkura N, et al. Foxp3 controls regulatory T-cell function by interacting with AML1/Runx1. Nature. 2007; 446(7136):685-689. (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. (Clone-specific: Flow cytometry). 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
562466 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.