Overview

  • Product name
    Anti-RUNX3 antibody - ChIP Grade
    See all RUNX3 primary antibodies
  • Description
    Rabbit polyclonal to RUNX3 - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ChIP, WB, EMSA, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein (Human).

  • Positive control
    • Raji nuclear extract. This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: A431

Properties

Applications

Our Abpromise guarantee covers the use of ab11905 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
ChIP Use at an assay dependent concentration. PubMed: 20714105
WB 1/500 - 1/1000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
EMSA Use at an assay dependent concentration.
ICC/IF 1/200.

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

  • Detection of AML2 by EMSA analysis. Nushift polyclonal antibody against AML2 factor creates a supershift in EMSA (lane 4).
  • ICC/IF image of ab11905 stained A431 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab11905 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Morita K  et al. RUNX transcription factors potentially control E-selectin expression in the bone marrow vascular niche in mice. Blood Adv 2:509-515 (2018). Read more (PubMed: 29500219) »
  • Zheng J  et al. DNA methylation affects metastasis of renal cancer and is associated with TGF-ß/RUNX3 inhibition. Cancer Cell Int 18:56 (2018). Read more (PubMed: 29651226) »
See all 10 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (RAJI cell lysate and HEK293T oveexpressing RUNX3)
Gel Running Conditions
Reduced Denaturing (10% SDS gel)
Loading amount
35 µg
Treatment
RUNX3 plasmid ytransfection for overexpresssion in 293T cells
Specification
RAJI cell lysate and HEK293T oveexpressing RUNX3
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Yupen Chung

Verified customer

Submitted Apr 25 2016

Answer

Thank you for your patience with this inquiry. I am very sorry for the delay.
We sourced this particular product externally and regrettably the laboratory cannot provide the information, which isoform of the RUNX3 was used to produce this antibody.
I am sorry that I cannot be of more help on this occasion. However, we can confirm that because the antibody was raised against the full length protein we would expect both isoforms to be detected with this antibody. Please notice that has not been tested specifically in the laboratory.
I wish you good luck with your further research. If you need more information please do not hesitate to contact us again.

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Question
Answer

Thank you for your phone call.

DISCOUNT CODE: xxx
Expiration date: xxx
Value: xxx

I am very pleased to hear you would like to accept our offer and test ab11905 in mouse. This code will give you xxx off your next order before the expiration date. To redeem this offer, please submit an Abreview for mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/abtrial.

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Answer

Thank you for taking the time to submit these comments. It looks to me from the gel that the gel has not run out properly. Could you please enquire with the customer as to how the gel was made and how long the running gel and stacking gel were left to polymerise for prior to use? Smeared banding such as that shown can be due to running the gel at too high a voltage. Since the gel can loose rigidity creating band smearing. Under what conditions was the gel ran? I suspect that reducing the voltage or running the gel in the cold room would reduce the observed smearing. I look forward to hearing from you again shortly.

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Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 210605 DESCRIPTION OF THE PROBLEM No specific signal, often nothing at all. SAMPLE Human T cells as well as PBMC depleted of monocytes and NK cells PRIMARY ANTIBODY ab11905, abcam, 1:1000. Incubation from 2 hr RT to overnight 4 degrees. 2 hr wash before addition of secondary. DETECTION METHOD ECL plus POSITIVE AND NEGATIVE CONTROLS USED Positive control: Jurkat lysate. No negative control ANTIBODY STORAGE CONDITIONS storage -20 degrees, aliquoted in two lots of 25 ?l SAMPLE PREPARATION lysed samples (buffer: 9 ml 5% triton x-100 in Tris HCL, 9g/l NaCl + 1 ml 0.5 M EDTA + complete protease inhibitor tablet (ROCHE)) mixed 1:1 with l?mmli buffer. AMOUNT OF PROTEIN LOADED Varied, sometimes different amounts of same sample used on 1 blot. ELECTROPHORESIS/GEL CONDITIONS 4-12% Nupage gel (Invitrogen), MOPS or MES buffer TRANSFER AND BLOCKING CONDITIONS Transfer 3 hrs, 4 degrees in transfer buffer (Invitrogen), 10% Methanol., blocked with 1 % BSA, 1 hr. SECONDARY ANTIBODY anti-rabbit-HRP, amaxa. 1:7000, 2 hr. wash two hr before development. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Incubation time and temperature, amount of sample, type of cell lysate, development time ADDITIONAL NOTES About purchase order number: The antibody was ordered by our technician, together with anti-RUNX1 (PON zumkehr210605. I have used the anti-RUNX1 antibody (ab11903) to satisfaction on the same samples and even blots as those used for this anti-RUNX3 antibody, also at a dilution of 1:1000 and with the same reagents. The much smaller amount of anti-RUNX3 supplied did not allow for dilutions less than 1:1000, without skyrocketing costs.

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Answer

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab11905. For Western blotting, we do suggest using this antibody at 1:500 - 1:1000 and so I do suggest trying ab11905 at 1:500 with incubation overnight at 4C. We also suggest using Raji nuclear extract as a positive control. I hope this helps. If you need additional assistance, then please contact us again.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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