Anti-RUNX3 antibody [R3-5G4] (ab40278)
Key features and details
- Mouse monoclonal [R3-5G4] to RUNX3
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-RUNX3 antibody [R3-5G4]
See all RUNX3 primary antibodies -
Description
Mouse monoclonal [R3-5G4] to RUNX3 -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment corresponding to Human RUNX3 aa 150 to the C-terminus (C terminal).
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Positive control
- ab40278 gave a positive result in the following whole cell lysates: Jurkat (Human T cell lymphoblast-like cell line) Raji (Human Burkitt's lymphoma cell line) SW480 (Human colon adenocarcinoma cell line)
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
R3-5G4 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab40278 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
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IHC-P | (1) |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa). |
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Function
CBF binds to the core site, 5'-PYGPYGGT-3', of a number of enhancers and promoters, including murine leukemia virus, polyomavirus enhancer, T-cell receptor enhancers, lck, IL-3 and GM-CSF promoters. -
Sequence similarities
Contains 1 Runt domain. -
Domain
A proline/serine/threonine rich region at the C-terminus is necessary for transcriptional activation of target genes. -
Post-translational
modificationsPhosphorylated on tyrosine residues by SRC. Phosphorylated by LCK and FYN. -
Cellular localization
Nucleus. Cytoplasm. The tyrosine phosphorylated form localizes to the cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 864 Human
- Omim: 600210 Human
- SwissProt: Q13761 Human
- Unigene: 170019 Human
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Alternative names
- Acute myeloid leukemia 2 protein antibody
- Acute myeloid leukemia gene 2 antibody
- AML 2 antibody
see all
Images
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: RUNX3 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)Lanes 1 - 3: Merged signal (red and green). Green - ab40278 observed at 44 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab40278 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. ab40278 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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IHC image of RUNX3 staining in human lymphoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab40278, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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Overlay histogram showing HEK293 cells stained with ab40278 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40278, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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All lanes : Anti-RUNX3 antibody [R3-5G4] (ab40278) at 1 µg/ml
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lane 3 :SW480 whole cell lysate (ab3957)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Rabbit Anti-Mouse IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 44 kDa
Additional bands at: 46 kDa (possible isoform)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (30)
ab40278 has been referenced in 30 publications.
- Douchi D et al. A Point Mutation R122C in RUNX3 Promotes the Expansion of Isthmus Stem Cells and Inhibits Their Differentiation in the Stomach. Cell Mol Gastroenterol Hepatol 13:1317-1345 (2022). PubMed: 35074568
- Lee SH et al. RUNX3 methylation drives hypoxia-induced cell proliferation and antiapoptosis in early tumorigenesis. Cell Death Differ 28:1251-1269 (2021). PubMed: 33116296
- Carrato C et al. Glioblastoma TCGA Mesenchymal and IGS 23 Tumors are Identifiable by IHC and have an Immune-phenotype Indicating a Potential Benefit from Immunotherapy. Clin Cancer Res 26:6600-6609 (2020). PubMed: 32998960
- Xiao Z et al. RUNX3 inhibits the invasion and migration of esophageal squamous cell carcinoma by reversing the epithelial-mesenchymal transition through TGF-ß/Smad signaling. Oncol Rep 43:1289-1299 (2020). PubMed: 32323849
- Liu H et al. Inhibition of PIM1 attenuates the stem cell-like traits of breast cancer cells by promoting RUNX3 nuclear retention. J Cell Mol Med 24:6308-6323 (2020). PubMed: 32307917