Anti-RUNX3 antibody - ChIP Grade [EPR20687] (ab224641)

Rabbit Recombinant Monoclonal RUNX3 antibody. Suitable for IP, ChIP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 10 publications.

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Images

ChIP - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (AB224641), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	ChIP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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Target data

See full target information RUNX3

Function

Forms the heterodimeric complex core-binding factor (CBF) with CBFB. RUNX members modulate the transcription of their target genes through recognizing the core consensus binding sequence 5'-TGTGGT-3', or very rarely, 5'-TGCGGT-3', within their regulatory regions via their runt domain, while CBFB is a non-DNA-binding regulatory subunit that allosterically enhances the sequence-specific DNA-binding capacity of RUNX. The heterodimers bind to the core site of a number of enhancers and promoters, including murine leukemia virus, polyomavirus enhancer, T-cell receptor enhancers, LCK, IL3 and GM-CSF promoters (By similarity). May be involved in the control of cellular proliferation and/or differentiation. In association with ZFHX3, up-regulates CDKN1A promoter activity following TGF-beta stimulation (PubMed:20599712). CBF complexes repress ZBTB7B transcription factor during cytotoxic (CD8+) T cell development. They bind to RUNX-binding sequence within the ZBTB7B locus acting as transcriptional silencer and allowing for cytotoxic T cell differentiation. CBF complexes binding to the transcriptional silencer is essential for recruitment of nuclear protein complexes that catalyze epigenetic modifications to establish epigenetic ZBTB7B silencing (By similarity).

Alternative names

AML2, CBFA3, PEBP2A3, RUNX3, Runt-related transcription factor 3, Acute myeloid leukemia 2 protein, Core-binding factor subunit alpha-3, Oncogene AML-2, Polyomavirus enhancer-binding protein 2 alpha C subunit, SL3-3 enhancer factor 1 alpha C subunit, SL3/AKV core-binding factor alpha C subunit, CBF-alpha-3, PEA2-alpha C, PEBP2-alpha C

Recommended products

Rabbit Recombinant Monoclonal RUNX3 antibody. Suitable for IP, ChIP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 10 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR20687
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

RUNX3 also known as Runt-related transcription factor 3 is a protein that plays an important role in regulating gene expression. The protein has a mass of approximately 44 kDa. RUNX3 is found mainly in tissues like the gastrointestinal tract and neural systems. It functions as a DNA-binding transcription factor that regulates various genes responsible for important cellular processes.

Biological function summary

The function of RUNX3 involves the regulation of cell proliferation differentiation and apoptosis. RUNX3 often forms a complex with the core-binding factor beta (CBFβ) enhancing its stability and DNA-binding capability. This protein is essential in maintaining cellular homeostasis and influencing the immune response particularly through its role in T cell development.

Pathways

RUNX3 is heavily involved in the TGF-beta signaling pathway and the Wnt signaling pathway. It interacts with several proteins such as SMADs in the TGF-beta pathway to modulate gene expression that affects cell growth and survival. Interaction with beta-catenin in the Wnt signaling pathway further emphasizes its importance in regulating cell fate decisions.

Associated diseases and disorders

Alterations in RUNX3 expression or function are linked to cancer including gastric cancer and colorectal cancer. It often shows reduced activity or expression in these cancers which associates with proteins like TGF-beta leading to impaired cell growth control. RUNX3 dysregulation is also connected to neurological disorders due to its role in neural development and function.

Product promise

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11 product images

ChIP - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Chromatin was prepared from U-937 (PMA treated or not) cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 5μg of ab224641 (red), and 20μl of protein A/G sepharose beads slurry (10μl of sepharose A beads + 10μl of sepharose G beads). 5μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Western blot - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Lanes 1 - 4: Merged signal (red and green). Green - ab224641 observed at 44-46 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.
ab224641 was shown to specifically react with RUNX3 in wild-type HAP1 cells as signal was lost in RUNX3 knockout cells. Wild-type and RUNX3 knockout samples were subjected to SDS-PAGE. ab224641 and Anti-Vinculin antibody [VIN-54] ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: RUNX3 knockout HAP1 whole cell lysate at 20 µg
Lane 3: Ramos whole cell lysate at 20 µg
Lane 4: HeLa whole cell lysate at 20 µg
Predicted band size: 44 kDa
Western blot - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Blocking/Dilution: 5% NFDM/TBST.
Negative control: MCF7 (PMID:21706051).
This target detects both predicted isoforms 44KDa and 46KDa that consistent with what has been described in the literature (PMID:23700080).
All lanes: Western blot - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641) at 1/1000 dilution
Lane 1: Raji (human Burkitt's lymphoma cell line) cell lysate at 20 µg
Lane 2: Ramos (human Burkitt's lymphoma cell line) cell lysate at 20 µg
Lane 3: MCF7 (human breast adenocarcinoma cell line) cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 44 kDa, 46 kDa
Exposure time: 5s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling RUNX3 with ab224641 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051), ready to use. Nuclear staining on lymphoid cells of human stomach (PMID:15514019; PMID:21786422) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051), ready to use.
Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunoprecipitation - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
RUNX3 was immunoprecipitated from 0.35 mg of Raji (human Burkitt's lymphoma cell line) whole cell lysate with ab224641 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224641 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution.
Lane 1: Raji whole cell lysate 10 μg (Input).
Lane 2: ab224641 IP in Raji whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab224641 in Raji whole cell lysate (-).
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Predicted band size: 44 kDa
Exposure time: 1s
Immunocytochemistry/ Immunofluorescence - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (human Burkitt's lymphoma cell line) cells labeling RUNX3 with ab224641 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on Ramos cell line. DAPI (blue) and anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution were used as counterstains.
The negative controls are as follows:
Negative control: MCF7 (human breast adenocarcinoma cell line) (PMID: 21706051).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor^®488 Goat anti-Rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution..
Western blot - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Blocking/Dilution: 5% NFDM/TBST.
This target detects both predicted isoforms 44KDa and 46KDa that consistent with what has been described in the literature (PMID:23700080).
All lanes: Western blot - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641) at 1/1000 dilution
All lanes: Human tonsil tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 44 kDa
Exposure time: 30s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling RUNX3 with ab224641 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051), ready to use. Nuclear staining on lymphoid cells of human gastric cancer (PMID:27566570) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051), ready to use.
Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunocytochemistry/ Immunofluorescence - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (human Burkitt's lymphoma cell line) cells labeling RUNX3 with ab224641 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on Raji cell line. DAPI (blue) and anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution were used as counterstains.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor^®488 Goat anti-Rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Immunohistochemical analysis of paraffin-embedded human diffuse large B cell lymphoma tissue labeling RUNX3 with ab224641 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051), ready to use. Nuclear staining on human diffuse large B cell lymphoma (PMID:27184221) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051), ready to use.
Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Flow Cytometry (Intracellular) - Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Raji (human Burkitt's lymphoma cell line) cell line labeling RUNX3 with ab224641 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.