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Alexa Fluor® 488 Rat Anti-Mouse Foxp3
Alexa Fluor® 488 Rat Anti-Mouse Foxp3
Multicolor flow cytometric analysis of Foxp3 expression in mouse splenocytes. Balb/c splenic leucocytes were 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 (Cat. No. 740007) and either Alexa Fluor® 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Plot) or Alexa Fluor® 488 Rat Anti-Mouse Foxp3 (Cat. No. 563487) antibodies at 0.5 µg/test. The bivariate pseudocolor density plot shows the correlated expression patterns of CD4 versus Foxp3 (or Ig Isotype control staining) for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis was performed using a BD™ LSR II Flow Cytometer System and FlowJo™ software.
Alexa Fluor® 488 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 Alexa Fluor® 647 Rat Anti-Mouse CD4 (Cat. No. 557681) and Alexa Fluor® 488 Rat Anti-Mouse Foxp3 (Cat. No. 563487) antibodies. The two-color flow cytometric dot plot shows the correlated expression patterns of CD4 versus Foxp3 for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of Foxp3 expression in mouse splenocytes. Balb/c splenic leucocytes were 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 (Cat. No. 740007) and either Alexa Fluor® 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; Left Plot) or Alexa Fluor® 488 Rat Anti-Mouse Foxp3 (Cat. No. 563487) antibodies at 0.5 µg/test. The bivariate pseudocolor density plot shows the correlated expression patterns of CD4 versus Foxp3 (or Ig Isotype control staining) for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis was performed using a BD™ LSR II Flow Cytometer System and FlowJo™ software.
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 Alexa Fluor® 647 Rat Anti-Mouse CD4 (Cat. No. 557681) and Alexa Fluor® 488 Rat Anti-Mouse Foxp3 (Cat. No. 563487) antibodies. The two-color flow cytometric dot plot shows the correlated expression patterns of CD4 versus Foxp3 for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry 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_2738236
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 to the dye under optimum conditions and unreacted dye was 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. 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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
563487 Rev. 2
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.

563487 Rev. 2
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
563487 Rev.2
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
563487 Rev. 2

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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.