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Flow cytometric analysis of EOMES expression in human peripheral blood lymphocytes. Peripheral blood mononuclear cells were stained intracellularly with PE Mouse Anti-Human CD8 antibody (Cat. No. 555367/557086/561949) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-EOMES antibody (Cat. No. 567166; Right Plot) at 0.5 µg/test using BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The bivariate pseudocolor density plot showing the correlated expression of EOMES (or Ig Isotype control staining) versus CD8 was 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™ 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 leucocytes. Spleen cells from C57BL/6 mice were stained intracellularly with PE Rat Anti-Mouse CD335 (NKp46) antibody (Cat.No. 560757) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Left Plot) or BD Horizon BV421 Mouse Anti-EOMES antibody (Right Plot) at 0.5 µg/test using BD Pharmingen™ Transcription Factor Buffer Set. The bivariate pseudocolor density plot showing the correlated expression of EOMES (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™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BV421 Mouse Anti-EOMES
BD Horizon™ BV421 Mouse Anti-EOMES
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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 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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Pacific Blue™ is a trademark of Life Technologies Corporation.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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 BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue™, and BD Horizon V450 cannot be used simultaneously.
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.