Overview

  • Product name

    Anti-RUNX3 antibody [2B3]
    See all RUNX3 primary antibodies
  • Description

    Mouse monoclonal [2B3] to RUNX3
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment, corresponding to amino acids 186-252 of Human RUNX3 expressed in E. Coli.

  • Positive control

    • Human RUNX3 recombinant protein; HEK293 cell lysate transfected with RUNX3 (aa186-252)-hIgGFc; NIH 3T3 cells; Human cervical cancer tissue.
  • General notes

    This product was changed from ascites to supernatant. Lot no’s high than GR206339-20 are from Tissue Culture Supernatant

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituent: 99% PBS

    0.5% protein stabilizer
  • Concentration information loading...
  • Purity

    Protein G purified
  • Purification notes

    Purified from tissue culture supernatant.
  • Clonality

    Monoclonal
  • Clone number

    2B3
  • Isotype

    IgG2b
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab135248 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
WB 1/500 - 1/2000. Predicted molecular weight: 44 kDa.
IHC-P 1/200 - 1/1000.
Flow Cyt 1/200 - 1/400.

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

ICC/IF 1/200 - 1/1000.

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

  • Immunohistochemical analysis of Paraffin-embedded Human cervical cancer tissue labelling RUNX3 with ab135248 at 1/200 dilution followed by DAB staining.
  • 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 - ab135248 observed at 44 kDa. Red - loading control, ab176560, observed at 50 kDa.

    ab135248 was shown to recognize RUNX3 in wild-type HAP1 cells as signal was lost at the expected MW in RUNX3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and RUNX3 knockout samples were subjected to SDS-PAGE. Ab135248 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1/500 dilution 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.

  • Flow cytometric analysis of NIH 3T3 cells labelling RUNX3 with ab135248 at 1/200 dilution (green). Negative control (red).
  • Anti-RUNX3 antibody [2B3] (ab135248) at 1/500 dilution + Recombinant Human RUNX3 protein

    Predicted band size: 44 kDa

  • All lanes : Anti-RUNX3 antibody [2B3] (ab135248) at 1/500 dilution

    Lane 1 : HEK293 cell lysate, non-transfected
    Lane 2 : HEK293 cell lysate, transfected with RUNX3 (amino acids 186-252)-hIgGFc

    Predicted band size: 44 kDa

References

This product has been referenced in:

  • Zhou WN  et al. Inactivation of RUNX3 protein expression in tongue squamous cell carcinoma and its association with clinicopathological characteristics. Mol Med Rep 19:885-894 (2019). Read more (PubMed: 30535462) »
  • Chen X  et al. Loss of expression rather than cytoplasmic mislocalization of RUNX3 predicts worse outcome in non-small cell lung cancer. Oncol Lett 15:5043-5055 (2018). IHC-P ; Human . Read more (PubMed: 29545901) »
See all 3 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Embryonic day 14.5 dorsal root ganglion)
Permeabilization
Yes - 0.1%Triton X-100
Specification
Embryonic day 14.5 dorsal root ganglion
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Formaldehyde

Dr. Frederic Clotman

Verified customer

Submitted Feb 09 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Thymus)
Permeabilization
Yes - Acetone
Specification
Thymus
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 25°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Oct 20 2015

Application
Flow Cytometry
Sample
Human Cell (293FT cell line)
Permeabilization
Yes - fixation/permeabilization concentrate- eBioscience 00-5123 (diluted 1:4 in diluent, eBioscience 00-5523)
Gating Strategy
Singlets were gated according to FSC-A on FSC-W
Specification
293FT cell line
Fixation
fixation/permeabilization concentrate- eBioscience 00-5123 (diluted 1:4 in diluent, eBioscience 00-5523)

Abcam user community

Verified customer

Submitted Sep 24 2015

Answer



Ich habe im Labor nachgefragt und freue mich Ihnen das Protokoll, das verwendet wurde um ab135248 zu testen, weiter zu leiten.



Flow Cytometry Protocol

1. Solutions and Reagents

1.1. 1X PBS

1.2. Blocking buffer: 0.5% BSA in 1X PBS

1.3. Filter buffer: 0.1%Triton-X100 and Blocking buffer

1.4. The in ice cold 4% paraformaldehyde (1% solution - optional for storing samples)

1.5. Fluorescently-conjugated secondary antibody (various forms)



2. Protocol

2.1. Collect 1x106 cells/sample.

2.2. Wash cells once with blocking buffer.

2.3. Fix cells with 4% paraformaldehyde and incubate at 4oC for 30 min.

2.4. Wash cells once with blocking buffer.

2.5. Add 0.5 ml filter buffer and incubate at 0 oC for 15 min.

2.6. Wash cells twice with blocking buffer.

2.7. Incubate cells in blocking buffer for 10 min at room temperature.

2.8. Add primary antibody at the appropriate dilution and incubate for 30 min at room temperature.

2.9. Wash twice with blocking buffer and incubate with fluorescently-conjugated secondary antibody for 30 min at room temperature.

2.10. Wash cells twice with blocking buffer.

2.11. Re-suspend cells in 1X PBS and analyze on flow cytometry. Samples can be kept in 1% paraformaldehyde at 4oC overnight.



3.1. Advice: Keep the cells in the dark on ice or at 4 oC in a fridge until your scheduled time for analysis. Analyze the cells on the flow cytometer as soon as possible.

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