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Alexa Fluor™ 647 Mouse Anti-EOMES
Alexa Fluor™ 647 Mouse Anti-EOMES
Flow cytometric analysis of EOMES expression in human peripheral blood lymphocytes. Peripheral blood mononuclear cells were stained intracellularly with BD Horizon™ BV421 Mouse Anti-Human CD8 antibody (Cat. No. 562428/562429) and either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; Left Plot) or Alexa Fluor™ 647 Mouse Anti-EOMES antibody (Cat. No. 567168; Right Plot) at 0.06 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.
Alexa Fluor™ 647 Mouse Anti-EOMES

Flow cytometric analysis of EOMES expression in mouse splenic leucocytes. Spleen cells from C57BL/6 mice were stained intracellularly with BD Horizon™ BV421 Rat Anti-Mouse CD335 (NKp46) antibody (Cat. No. 562850) and either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Left Plot) or Alexa Fluor™ 647 Mouse Anti-EOMES antibody (Right Plot) at 0.06 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.

Flow cytometric analysis of EOMES expression in human peripheral blood lymphocytes. Peripheral blood mononuclear cells were stained intracellularly with BD Horizon™ BV421 Mouse Anti-Human CD8 antibody (Cat. No. 562428/562429) and either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; Left Plot) or Alexa Fluor™ 647 Mouse Anti-EOMES antibody (Cat. No. 567168; Right Plot) at 0.06 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 BD Horizon™ BV421 Rat Anti-Mouse CD335 (NKp46) antibody (Cat. No. 562850) and either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Left Plot) or Alexa Fluor™ 647 Mouse Anti-EOMES antibody (Right Plot) at 0.06 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.

Product Details
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BD Pharmingen™
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 ≤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 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 cell and CompBead to ensure that BD Comp beads 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. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  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.
567168 Rev. 1
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.

        

567168 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
567168 Rev.1
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
567168 Rev. 1

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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.