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BV421 Rat Anti-Mouse Foxp3
BV421 Rat Anti-Mouse Foxp3
Multicolor flow cytometric analysis of Foxp3 expression in BALB/c mouse splenocytes. Splenic leucocytes from BALB/c mice were stained with APC Rat anti-Mouse CD4 antibody (Cat. No. 553051/561091), and fixed and permeabilized using working solutions of the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with BD Horizon™ BV421Rat Anti-Mouse Foxp3 antibody (Cat. No. 562996). 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 leucocytes. Flow cytometry was performed using a BD LSRFortessa™ Cell Analyzer System.
Multicolor flow cytometric analysis of Foxp3 expression in BALB/c mouse splenocytes. Splenic leucocytes from BALB/c mice were stained with APC Rat anti-Mouse CD4 antibody (Cat. No. 553051/561091), and fixed and permeabilized using working solutions of the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with BD Horizon™ BV421Rat Anti-Mouse Foxp3 antibody (Cat. No. 562996). 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 leucocytes. Flow cytometry was performed using a BD LSRFortessa™ Cell Analyzer 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_2737940
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.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  9. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  11. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
562996 Rev. 2
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

562996 Rev. 2
Format Details
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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.
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BV421
Violet 405 nm
407 nm
423 nm
562996 Rev.2
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. (Clone-specific: Flow cytometry). 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
562996 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.