Two-color flow cytometric analysis of EOMES expression in Human peripheral blood lymphocytes. Peripheral blood mononuclear cells (PMBC) were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with APC Mouse Anti-Human CD8 antibody (Cat. No. 561953/555369) and with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; Left Plot) or BD Horizon™ RB613 Mouse Anti-EOMES antibody (Cat. No. 571362/571363; Right Plot) at 0.5 µg/test. The bivariate pseudocolor density plot showing EOMES (or Ig Isotype control staining) versus CD8 was derived from gated 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.
Two-color flow cytometric analysis of EOMES expression on Mouse splenic leukocytes. C57BL/6 Mouse splenocytes were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD335 antibody (Cat. No. 562850) and with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Left Plot) or BD Horizon™ RB613 Mouse Anti-EOMES antibody (Right Plot) at 0.5 µg/test using BD Pharmingen™ Transcription Factor Buffer Set. The bivariate pseudocolor density plot showing EOMES (or Ig Isotype control staining) versus CD335 was derived from gated events with the forward and side light-scatter characteristics of intact splenic leukocytes. 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.
Two-color flow cytometric analysis of EOMES expression in Human peripheral blood lymphocytes. Peripheral blood mononuclear cells (PMBC) were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with APC Mouse Anti-Human CD8 antibody (Cat. No. 561953/555369) and with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; Left Plot) or BD Horizon™ RB613 Mouse Anti-EOMES antibody (Cat. No. 571362/571363; Right Plot) at 0.5 µg/test. The bivariate pseudocolor density plot showing EOMES (or Ig Isotype control staining) versus CD8 was derived from gated 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.
Two-color flow cytometric analysis of EOMES expression on Mouse splenic leukocytes. C57BL/6 Mouse splenocytes were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD335 antibody (Cat. No. 562850) and with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Left Plot) or BD Horizon™ RB613 Mouse Anti-EOMES antibody (Right Plot) at 0.5 µg/test using BD Pharmingen™ Transcription Factor Buffer Set. The bivariate pseudocolor density plot showing EOMES (or Ig Isotype control staining) versus CD335 was derived from gated events with the forward and side light-scatter characteristics of intact splenic leukocytes. 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
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
8320, 13813
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- CF™ is a trademark of Biotium, Inc.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
RB613 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
571106
APC Mouse Anti-Human CD8 RUO
Size
500 Tests
Cat No.
561953
571363 Rev. 1
Antibody Details
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.
571363 Rev. 1
Format Details
RB613
The BD Horizon RealBlue™ 613 (RB613) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492-nm and an emission maximum (Em Max) at 613-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB613 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with reduced excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB613 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-CF594. RB613 is on average brighter than PE-CF594 off the blue laser.
RB613
Blue 488 nm
492 nm
613 nm
571363 Rev.1
Citations & References
View product citations for antibody "571363" on CiteAb
Development References (6)
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Lee J, Nicosia M, Hong ES, et al. Sex-Biased T-cell Exhaustion Drives Differential Immune Responses in Glioblastoma.. Cancer Discov. 2023; 13(9):2090-2105. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
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Morris AB, Farley CR, Pinelli DF, et al. Signaling through the Inhibitory Fc Receptor FcγRIIB Induces CD8+ T Cell Apoptosis to Limit T Cell Immunity.. Immunity. 2020; 52(1):136-150.e6. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
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Noack F, Vangelisti S, Raffl G, et al. Multimodal profiling of the transcriptional regulatory landscape of the developing mouse cortex identifies Neurog2 as a key epigenome remodeler.. Nat Neurosci. 2022; 25(2):154-167. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
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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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Reuter MA, Pombo C, Betts MR. Cytokine production and dysregulation in HIV pathogenesis: lessons for development of therapeutics and vaccines.. Cytokine Growth Factor Rev. 23(4-5):181-91. (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
571363 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.