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BUV395 Mouse Anti-EOMES
BUV395 Mouse Anti-EOMES
Flow cytometric analysis of EOMES expression in human peripheral blood lymphocyte populations. Peripheral blood mononuclear cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). Cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD8 antibody (Cat. No. 562428/ 562429) and either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Plot) or BD Horizon™ BUV395 Mouse Anti-EOMES antibody (Cat. No. 566749; Right Plot) at 1 µg/test. The bivariate pseudocolor density plots showing EOMES expression (or Ig isotype control staining) versus CD8 were derived from events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data 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.
BUV395 Mouse Anti-EOMES

Flow cytometric analysis of EOMES expression in mouse splenic leucocyte populations. Spleen cells from C57BL/6 mice were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set. Cells were stained intracellularly with Alexa Fluor® 647 Rat anti-mouse CD335 (NKp46) antibody (Cat.No. 560755) and either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Left Plot) or BD Horizon™ BUV395 Mouse Anti-EOMES antibody (Right Plot) at 1.0 µg/test. The bivariate pseudocolor density plots showing EOMES expression (or Ig isotype control staining) versus CD355 (NKp46) were derived from events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data 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.

Flow cytometric analysis of EOMES expression in human peripheral blood lymphocyte populations. Peripheral blood mononuclear cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). Cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD8 antibody (Cat. No. 562428/ 562429) and either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Plot) or BD Horizon™ BUV395 Mouse Anti-EOMES antibody (Cat. No. 566749; Right Plot) at 1 µg/test. The bivariate pseudocolor density plots showing EOMES expression (or Ig isotype control staining) versus CD8 were derived from events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data 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.

Flow cytometric analysis of EOMES expression in mouse splenic leucocyte populations. Spleen cells from C57BL/6 mice were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set. Cells were stained intracellularly with Alexa Fluor® 647 Rat anti-mouse CD335 (NKp46) antibody (Cat.No. 560755) and either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Left Plot) or BD Horizon™ BUV395 Mouse Anti-EOMES antibody (Right Plot) at 1.0 µg/test. The bivariate pseudocolor density plots showing EOMES expression (or Ig isotype control staining) versus CD355 (NKp46) were derived from events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data 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
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BD Horizon™
C77258; T-box brain protein 2; TBR-2; TBR2; Tbr2; eomesodermin homolog
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG1, κ
Human EOMES aa 1-275 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
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 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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Antibody Details
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X4-83

The X4-83 monoclonal antibody specifically binds to human and mouse Eomesodermin (EOMES), a transcription factor of the T-box family that is also known as T-box brain 2 (TBR2). The aligned sequences of human and mouse EOMES proteins are 86.7% identical and their DNA binding T-box domains are about 74% identical to those of the human and mouse T-bet transcription factors. The name Eomesodermin comes from the Greek word "eo", meaning dawn, and "mesoderm" because it was first identified to be essential for early stages of mesoderm formation. Later in embryonic development, the EOMES transcription factor is important in the differentiation of neurons. EOMES and T-bet are the only T-box transcription factors that are expressed in the immune system, and some of their functions appear to be redundant. EOMES is expressed in cytotoxic T lymphocytes and NK cells, and at lower levels in T helper lymphocytes. EOMES is one of several transcription factors that control the differentiation and survival of effector and memory CD8-positive T lymphocytes, NK cells, and ILC1.

The antibody was conjugated to BD Horizon BUV395 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. With an Ex Max near 348 nm and an Em Max near 395 nm, BD Horizon BUV395 can be excited by the ultraviolet laser (355 nm) laser and detected with a 379/28 filter. This dye has been exclusively developed by BD Biosciences as an optimal dye for use on instruments equipped with the ultraviolet laser and has virtually no spillover into any other detector.

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Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
Ultraviolet 355 nm
348 nm
395 nm
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Citations & References
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Development References (2)

  1. Picozzi P, Wang F, Cronk K, Ryan K. Eomesodermin requires transforming growth factor-beta/activin signaling and binds Smad2 to activate mesodermal genes. J Biol Chem. 2009; 284(4):2397-408. (Biology). View Reference
  2. Zhang J, Marotel M, Fauteux-Daniel S, et al. T-bet and Eomes govern differentiation and function of mouse and human NK cells and ILC1. Eur J Immunol. 2018; 48(5):738-750. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.