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BV421 Mouse Anti-RUNX3
BV421 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), FITC Mouse Anti-Human CD3 (Cat. No. 555332/561806/561807) and APC Mouse Anti-Human CD56 (Cat. No. 555518) 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™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438) or BD Horizon™BV421 Mouse Anti-RUNX3 antibody (Cat. No. 565742). 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 APC Rat Anti-Mouse CD4 antibody (Cat. No. 553048/553049/561837) and then fixed and permeabilized (as described above), followed by staining with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Upper Plot) or BD Horizon™ BV421 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 cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System.
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), FITC Mouse Anti-Human CD3 (Cat. No. 555332/561806/561807) and APC Mouse Anti-Human CD56 (Cat. No. 555518) 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™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438) or BD Horizon™BV421 Mouse Anti-RUNX3 antibody (Cat. No. 565742). 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 APC Rat Anti-Mouse CD4 antibody (Cat. No. 553048/553049/561837) and then fixed and permeabilized (as described above), followed by staining with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Upper Plot) or BD Horizon™ BV421 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 cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System.
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 BSA and ≤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 unconjugated antibody and free dye were 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 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

  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Pacific Blue™ is a trademark of Life Technologies Corporation.
  10. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565742 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 BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

565742 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
565742 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
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565742 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.