Anti-RUNX3 antibody [R3-5G4] (ab40278)

Knockout Tested Mouse monoclonal RUNX3 antibody [R3-5G4]. Validated in WB, IP, IHC, EMSA, Flow Cyt and tested in Human. Cited in 18 publication(s). Independently reviewed in 2 review(s).

Overview

  • 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, IP, WB, EMSA, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment corresponding to Human RUNX3 aa 150 to the C-terminus (C terminal).

  • 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)
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

  • 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...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    R3-5G4
  • Isotype

    IgG1
  • Research areas

Applications

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
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
EMSA Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • 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
    modifications

    Phosphorylated 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
  • Database links

  • Alternative names

    • Acute myeloid leukemia 2 protein antibody
    • Acute myeloid leukemia gene 2 antibody
    • AML 2 antibody
    • AML2 antibody
    • CBF alpha 3 antibody
    • CBF-alpha-3 antibody
    • CBFA 3 antibody
    • CBFA3 antibody
    • Core binding factor alpha 3 subunit antibody
    • core binding factor antibody
    • Core binding factor runt domain alpha subunit 3 antibody
    • Core binding factor subunit alpha 3 antibody
    • core-binding factor antibody
    • Core-binding factor subunit alpha-3 antibody
    • Oncogene AML 2 antibody
    • Oncogene AML-2 antibody
    • PEA2 alpha C antibody
    • PEA2-alpha C antibody
    • PEBP2 alpha C antibody
    • PEBP2-alpha C antibody
    • Pebp2a3 antibody
    • PEBP2aC antibody
    • Polyomavirus enhancer binding protein 2 alpha C subunit antibody
    • Polyomavirus enhancer-binding protein 2 alpha C subunit antibody
    • runt domain alpha subunit 3 antibody
    • runt related transcription factor 3 antibody
    • Runt-related transcription factor 3 antibody
    • RUNX 3 antibody
    • Runx3 antibody
    • RUNX3_HUMAN antibody
    • SL3 3 enhancer factor 1 alpha C subunit antibody
    • SL3-3 enhancer factor 1 alpha C subunit antibody
    • SL3/AKV core binding factor alpha C subunit antibody
    • SL3/AKV core-binding factor alpha C subunit antibody
    • Transcription factor AML2 antibody
    see all

Images

  • 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.

  • 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.
  • 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.

  • 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)

References

This product has been referenced in:

  • Choo SY  et al. Runx3 inhibits endothelial progenitor cell differentiation and function via suppression of HIF-1a activity. Int J Oncol 54:1327-1336 (2019). Read more (PubMed: 30968151) »
  • Lee JW  et al. RUNX3 regulates cell cycle-dependent chromatin dynamics by functioning as a pioneer factor of the restriction-point. Nat Commun 10:1897 (2019). Read more (PubMed: 31015486) »
See all 20 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

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Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (stomach)
Specification
stomach
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA (pH 9.0)
Permeabilization
No
Blocking step
RE7102, NovoLink Polymer Detection System as blocking agent for 5 minute(s) · Concentration: 1% · Temperature: RT°C

Dr. Yuan LI

Verified customer

Submitted Aug 20 2010

Application
ChIP
Sample
Human Cell lysate - whole cell (purified CD34+ cells)
Specification
purified CD34+ cells
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 5 minute(s) and 0 second(s)
Specification of the cross-linking agent: 0.4% formaldehyde
Detection step
Real-time PCR
Positive control
known bound region
Negative control
known neg control region

Abcam user community

Verified customer

Submitted Oct 06 2009

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