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R718 Mouse Anti-RUNX3
R718 Mouse Anti-RUNX3

Multicolor flow cytometric analysis of RUNX3 expression in human peripheral blood cells and mouse lymph node cells.

Panel 1. Human peripheral blood mononuclear cells (PBMC) were stained with BD Horizon™ BUV395 Mouse Anti-Human CD8α (Cat. No. 563796), PE Mouse Anti-Human CD3 (Cat. No. 555333/561808/561809) and FITC Mouse Anti-Human CD56 (NCAM-1) (Cat. No. 562794) antibodies. The cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928) or BD Horizon™ R718 Mouse Anti-RUNX3 antibody (Cat. No. 565741). The bivariate pseudocolor density plots on the left show the correlated expression of Ig Isotype control staining (Upper Left Plot) or RUNX3 (Bottom Left Plot) versus CD8α for gated events with the light scattering characteristics of intact lymphocytes. Intact cells were also gated as indicated (Upper Right Plot) and used for the generation of the overlaid dot plot figure (Bottom Right Plot) showing the correlated expression of RUNX3 versus CD3 for the CD8+CD56+ (blue), CD8+CD56- (green), and CD8-CD56+ (red) cell subsets.

Panel 2. C57BL/6 mouse lymph node cells (LNC) were stained with FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553047/553046/561835) and then fixed and permeabilized (as described above), followed by staining with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Upper Plot) or BD Horizon™ R718 Mouse Anti-RUNX3 antibody (Bottom Plot). The bivariate pseudocolor density plots showing the correlated expression of RUNX3 (or Ig Isotype control staining) versus CD4 were derived from gated events with the light-scatter characteristics of intact lymphocytes.

Flow cytometry and data analysis was performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ software.

Multicolor flow cytometric analysis of RUNX3 expression in human peripheral blood cells and mouse lymph node cells.

Panel 1. Human peripheral blood mononuclear cells (PBMC) were stained with BD Horizon™ BUV395 Mouse Anti-Human CD8α (Cat. No. 563796), PE Mouse Anti-Human CD3 (Cat. No. 555333/561808/561809) and FITC Mouse Anti-Human CD56 (NCAM-1) (Cat. No. 562794) antibodies. The cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928) or BD Horizon™ R718 Mouse Anti-RUNX3 antibody (Cat. No. 565741). The bivariate pseudocolor density plots on the left show the correlated expression of Ig Isotype control staining (Upper Left Plot) or RUNX3 (Bottom Left Plot) versus CD8α for gated events with the light scattering characteristics of intact lymphocytes. Intact cells were also gated as indicated (Upper Right Plot) and used for the generation of the overlaid dot plot figure (Bottom Right Plot) showing the correlated expression of RUNX3 versus CD3 for the CD8+CD56+ (blue), CD8+CD56- (green), and CD8-CD56+ (red) cell subsets.

Panel 2. C57BL/6 mouse lymph node cells (LNC) were stained with FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553047/553046/561835) and then fixed and permeabilized (as described above), followed by staining with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Upper Plot) or BD Horizon™ R718 Mouse Anti-RUNX3 antibody (Bottom Plot). The bivariate pseudocolor density plots showing the correlated expression of RUNX3 (or Ig Isotype control staining) versus CD4 were derived from gated events with the light-scatter characteristics of intact lymphocytes.

Flow cytometry and data analysis was performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ software.

Product Details
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BD Horizon™
AML2; AML-2; CBFA3; PEBP2aC; PEBP2A3
Human (QC Testing), Mouse (Tested in Development)
Mouse BALB/c IgG1, κ
Human RUNX3 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
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 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

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
567641 Rev. 1
Antibody Details
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R3-5G4

The R3-5G4 monoclonal antibody specifically binds to Runt-related transcription factor 3 (RUNX3) which is also known as Acute myeloid leukemia 2 protein (AML2), Core-binding factor subunit alpha-3 (CBFA3), and Polyomavirus enhancer-binding protein 2 alpha C subunit (PEBP2aC).  RUNX3 is a member of the RUNX transcription factor family which includes RUNX1-3. RUNX3 is a key regulator of gene expression related to the development and differentiation of cells within the nervous and immune systems. In the immune system, RUNX3 is particularly involved in the commitment of CD8+ T cells in the thymus. RUNX3 is highly expressed and essential for the cytotoxic functions of NK cells, peripheral CD8+T cells and CD4+CD8αα intraepithelial lymphocytes in the gut. RUNX3 also plays a role in the differentiation and effector functions of Th1 cells. RUNX3 is activated downstream of the TGF-β signaling pathway and can play a role in tumor suppression. Aberrant expression of RUNX3 has been associated with tumorigenesis including the development of gastric and other cancers.

The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.

567641 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
R718
Red 627-640 nm
695 nm
718 nm
567641 Rev.1
Citations & References
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Development References (6)

  1. Chuang LS, Ito K, Ito Y. RUNX family: Regulation and diversification of roles through interacting proteins. Int J Cancer. 2013; 132(6):1260-1271. (Biology). View Reference
  2. Djuretic IM, Cruz-Guilloty F, Rao A. Regulation of gene expression in peripheral T cells by Runx transcription factors. Adv Immunol. 2009; 104(1):23. (Biology). View Reference
  3. Ito K, Liu Q, Salto-Tellez M, et al. RUNX3, a novel tumor suppressor, is frequently inactivated in gastric cancer by protein mislocalization. Cancer Res. 2005; 65(17):7743-7750. (Immunogen: Western blot). View Reference
  4. Lotem J, Levanon D, Negreanu V, Leshkowitz D, Friedlander G, Groner Y. Runx3-mediated transcriptional program in cytotoxic lymphocytes. PLoS ONE. 2013; 8(11):e80467. (Biology). View Reference
  5. Reis BS, Rogoz A, Costa-Pinto FA, Taniuchi I, Mucida D. Mutual expression of the transcription factors Runx3 and ThPOK regulates intestinal CD4(+) T cell immunity. Nat Immunol. 2013; 14(3):271-280. (Biology). View Reference
  6. Setoguchi R, Tachibana M, Naoe Y, et al. Repression of the transcription factor Th-POK by Runx complexes in cytotoxic T cell devel. Science. 2008; 19(5864):822-825. (Biology). View Reference
View All (6) View Less
567641 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products 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.

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.