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Multicolor flow cytometric analysis of EOMES expression in human peripheral blood lymphocytes. 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™ BUV737 Mouse IgG1, κ Isotype Control (Cat. No. 612758; Left Plot) or BD Horizon™ BUV737 Mouse Anti-EOMES antibody (Cat. No. 567170; 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.
Multicolor 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 BD Horizon™ BV421 Rat anti-mouse CD335 (NKp46) antibody (Cat.No. 564065) and either BD Horizon™ BUV737 Mouse IgG1, κ Isotype Control (Left Plot) or BD Horizon™ BUV737 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.
BD Horizon™ BUV737 Mouse Anti-EOMES
BD Horizon™ BUV737 Mouse Anti-EOMES
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Preparation And Storage
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 CompBead 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 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).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
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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 BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 737 nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 nm filter. Due to the excitation of the acceptor dye by the red laser line, there may be significant spillover into red laser detectors with filters in the 700-720 nm range.
Development References (2)
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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
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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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.
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