Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Simple Western, CyTOF-ready

Cited:

Western Blot, Flow Cytometry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 527327
View all RUNX3/CBFA3 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human RUNX3/CBFA3
Lys186-Tyr415
Accession # Q13761

Specificity

Detects human and mouse RUNX3/CBFA3 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human/Mouse RUNX3/CBFA3 Antibody

Detection of RUNX3/CBFA3 antibody in Human PBMC antibody by Flow Cytometry.

Detection of RUNX3/CBFA3 in Human PBMC by Flow Cytometry.

Human PBMC unstimulated (light orange filled histogram) or treated with 50ng/mL PMA (dark orange filled histogram) were stained with Mouse Anti-Human/Mouse RUNX3/CBFA3 Monoclonal Antibody (Catalog # MAB3765) or isotype control antibody (Catalog # MAB003, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
RUNX3/CBFA3 antibody in Human PBMCs by Immunocytochemistry (ICC).

RUNX3/CBFA3 in Human PBMCs.

RUNX3/CBFA3 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human/Mouse RUNX3/CBFA3 Monoclonal Antibody (Catalog # MAB3765) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Human and Mouse RUNX3/CBFA3 antibody by Western Blot.

Detection of Human and Mouse RUNX3/CBFA3 by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line, Daudi human Burkitt's lymphoma cell line, and DA3 mouse myeloma cell line. Gels were loaded with 30 µg of whole cell lysate (WCL), 20 µg of cytoplasmic (Cyto), and 10 µg of nuclear extracts (Nuc). PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse RUNX3/CBFA3 Monoclonal Antibody (Catalog # MAB3765) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Specific bands were detected for RUNX3/CBFA3 at approximately 48 and 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human RUNX3/CBFA3 antibody by Simple WesternTM.

Detection of Human RUNX3/CBFA3 by Simple WesternTM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and Daudi human Burkitt's lymphoma cell line, loaded at 0.5 mg/mL. A specific band was detected for RUNX3/CBFA3 at approximately 54 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse RUNX3/CBFA3 Monoclonal Antibody (Catalog # MAB3765). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of RUNX3/CBFA3 by Flow Cytometry

Detection of RUNX3/CBFA3 by Flow Cytometry

RA and RAR alpha reciprocally regulate the expression of the positive and negative LC-regulating transcription factors, Runx3 and C/EBP beta. a Impact of RA and RAR alpha deficiency on Runx3 expression at mRNA level. b Impact of RA and RAR alpha deficiency on Runx3 expression at protein level. c Effect of enforced Runx3 expression on BM-derived LC differentiation in the presence and absence of RA. The data shown are gated for transduced Thy1.1+CD11c+ cells. d Impact of RA and RAR alpha deficiency on expression of Cebpb mRNA. e Impact of RA and RAR alpha deficiency on expression of C/EBP beta protein. f Effect of dnC/EBP beta on BM-derived LC differentiation in the presence and absence of RA. BM cells from WT or ∆RaraCD11c mice were cultured with GM-CSF and TGF beta 1 for 5 days (3 days following retroviral transduction) in the presence of At-RA (10 nM except in panels c and f where 0.1 nM was used) in a medium containing charcoal-treated FBS. Representative and combined data (n = 3–7) from at least 3 experiments are shown. Significant differences from controls by one-way ANOVA with Bonferroni adjustments (p < 0.05)* or between indicated groups by two-way ANOVA with Tukey adjustments (p < 0.05)**. Error bars are defined as s.e.m Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30254197), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of RUNX3/CBFA3 by Flow Cytometry

Detection of RUNX3/CBFA3 by Flow Cytometry

RA regulates CEBPb and RUNX3 expression and human langerin+ cell differentiation from blood monocytes. a Human LC differentiation from blood monocytes in regular vs. charcoal FBS. b At-RA promotes C/EBP beta + non-LC differentiation. c RA and RAR alpha antagonists reciprocally regulate human LC differentiation. d Effects of At-RA and BMS493 on the expression of human CEBPb and RUNX3. Human blood CD14+ monocytes were cultured in GM-CSF and TGF-beta 1 for 5–7 days in media containing charcoal FBS except in panel A where regular FBS was also used. At-RA was used at 1 nM, and BMS493 and Ro4153 were used at 500 nM. Representative and combined data (n = 5–7) from at least 5 experiments are shown. *Significant differences from control by Mann–Whitney U test (p < 0.05, unpaired, 2-sided). Error bars are defined as s.e.m. Langerhans cells (LC) and langerin-expressing conventional dendritic cells are made from distinct progenitors and enriched in the distinct microenvironments of the skin. Here the authors show that these immune cells are regulated by RAR alpha via simultaneous induction of LC-promoting Runx3 and repression of LC-inhibiting C/EBP beta Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30254197), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse RUNX3/CBFA3 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs) and DA3 mouse myeloma cell line

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: Human PBMC treated with PMA and mouse splenocytes fixed with paraformaldehyde and permeabilized with saponin

Simple Western

10 µg/mL
Sample: Jurkat human acute T cell leukemia cell line and Daudi human Burkitt's lymphoma cell line

Western Blot

1 µg/mL
Sample: Jurkat human acute T cell leukemia cell line, Daudi human Burkitt's lymphoma cell line, and DA3 mouse myeloma cell line

Reviewed Applications

Read 1 review rated 5 using MAB3765 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: RUNX3/CBFA3

RUNX3, also called CBFA3, AML-2 or PEBP2-alpha C, is a member of the Runt domain family of nuclear transcriptional regulators. All of the RUNX proteins form dimers with CBF-beta. The runt domain (aa 54-186) is required for DNA binding, while a pro/ser/thr-rich region (aa 191-415) transcriptionally activates target genes. Isoform 2 has an alternate 19 aa in place of the N-terminal 5 aa of isoform 1. The 415 aa Human RUNX3 shares 91% aa identity with mouse or rat RUNX3. RUNX3 is necessary for growth control of gastric epithelium, neurogenesis of dorsal root ganglia, and T cell differentiation. RUNX3 expression is frequently mutated in tumors and appears to be silenced by methylation.

Long Name

Runt-related Transcription Factor 3

Alternate Names

AML2, CBFA3, PEBP2A3, PEBP2AC

Entrez Gene IDs

864 (Human); 12399 (Mouse); 156726 (Rat)

Gene Symbol

RUNX3

UniProt

Q13761

Additional RUNX3/CBFA3 Products

Product Documents for Human/Mouse RUNX3/CBFA3 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse RUNX3/CBFA3 Antibody

For research use only

Citations for Human/Mouse RUNX3/CBFA3 Antibody

Customer Reviews for Human/Mouse RUNX3/CBFA3 Antibody (1)

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  • Human/Mouse RUNX3/CBFA3 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Peripheral blood mononuclear cells (PBMCs)
    Species: Mouse
    Verified Customer | Posted 12/18/2021
    Human/Mouse RUNX3/CBFA3 Antibody MAB3765

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