Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20687] to RUNX3 - BSA and Azide free
  • Suitable for: ChIP, IP, Flow Cyt, IHC-P, WB, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-RUNX3 antibody [EPR20687] - BSA and Azide free
    See all RUNX3 primary antibodies

  • Description

    Rabbit monoclonal [EPR20687] to RUNX3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC: Human stomach tissue.

  • General notes

    Ab227125 is the carrier-free version of ab224641. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab227125 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 production
    For more information see here.

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

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    pH: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR20687

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    Western blot - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    All lanes : Anti-RUNX3 antibody [EPR20687] - ChIP Grade (ab224641) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : RUNX3 knockout HAP1 whole cell lysate
    Lane 3 : Ramos whole cell lysate
    Lane 4 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 44 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab224641 observed at 44-46 kDa. Red - loading control, 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 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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224641).

  • Immunoprecipitation - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    Immunoprecipitation - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

    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) (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 (ab172730) instead of ab224641 in Raji whole cell lysate (-).

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224641).

  • Flow Cytometry - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    Flow Cytometry - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

    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 (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) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224641).

  • Immunocytochemistry/ Immunofluorescence - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    Immunocytochemistry/ Immunofluorescence - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

    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) (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) (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 (ab150077) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224641).

  • Immunocytochemistry/ Immunofluorescence - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    Immunocytochemistry/ Immunofluorescence - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

    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) (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) (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 (ab150077) at 1/1000 dilution..

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224641).

  • ChIP - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    ChIP - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224641).

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

    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) (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) (ab97051), ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224641).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

    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) (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) (ab97051), ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224641).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

    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) (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) (ab97051), ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab224641).

  • Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)
    Anti-RUNX3 antibody [EPR20687] - BSA and Azide free (ab227125)

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