Recombinant
RabMAb

Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336)

Overview

  • Product name
    Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099]
    See all RUNX1 / AML1+RUNX3+RUNX2 primary antibodies
  • Description
    Rabbit monoclonal [EPR3099] to RUNX1 / AML1 + RUNX3 + RUNX2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human RUNX1/ AML1 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab177141)

  • Positive control
    • WB: MOLT4, WEHI-3, CTLL-2 and Raw264.7 cell lysate; mouse and rat thymus tissue lysate, mouse spleen tissue lysate and fetal thymus tissue lysate. IHC: Human tonsil tissue. IP: Molt-4 cell lysate
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab92336 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 1/5000 - 1/10000. Predicted molecular weight: 49 kDa.Can be blocked with RUNX1 / AML1 peptide (ab177141).
IP 1/50.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

The use of an HRP/AP polymerized secondary antibody will give a stronger signal.

Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100 - 1/250.

Target

Images

  • All lanes : Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1.28 µg/ml (purified)

    Lane 1 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 2 : Molt-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate
    Lane 3 : WEHI-3 (Mouse leukemia lymphoblast) whole cell lysate
    Lane 4 : Mouse thymus lysate
    Lane 5 : CTLL-2 (Mouse T lymphocyte) whole cell lysate
    Lane 6 : Rat thymus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/ml

    Predicted band size: 49 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM /TBST 

  • ab92336 (purified) at 1/500 immunoprecipitating RUNX1 / AML1 + RUNX3 + RUNX2 in 10 μg Molt-4 (Human lymphoblastic leukemia T lymphoblast)whole cell lysate (Lanes 1 and 2, observed at 49 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730) instead of ab92336 in Molt-4 whole cell lysate. For western blotting, HRP Veriblot for IP (ab131366) was used as the secondary antibody (1/1000).

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

  • All lanes : Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1.28 µg/ml (purified)

    Lane 1 : Mouse spleen lysate
    Lane 2 : Mouse thymus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg

    Predicted band size: 49 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM /TBST

  • RUNX2 recombinant protein full length, with N-terminal HIS tag, expressed in E.Coli.

    RUNX3 overexpression and empty vector control lysates created in HEK293T cells.  The protein contains a C-terminal DDK tag.

  • All lanes : Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1/10000 dilution

    Lane 1 : MOLT4
    cell lysate
    Lane 2 : fetal thymus lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

    Predicted band size: 49 kDa

  • Immunohistochemistry staining of RUNX1 / AML1 in formalin-fixed, paraffin-embedded Human tonsil tissue using 1/100 ab92336.
  • Flow cytometric analysis of permeabilized Molt-4 cells using anti-RUNX1 ab92336 (red) or a rabbit IgG (negative) (green).
  • ab92336 staining RUNX1 / AML1 in human glioblastoma cell line by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab92336 at a 1/50 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/400 dilution. Nuclei are counterstained with DAPI.

    See Abreview

  • ab92336 staining RUNX1 / AML1 in rat glioblastoma cell line C6 by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab92336 at a 1/50 dilution for 16 hours at 4°C. The secondary used was a Cy3 conjugated goat anti-rabbit polyclonal used at a 1/400 dilution. Nuclei are counterstained with DAPI.

    See Abreview

References

This product has been referenced in:
  • Draper JE  et al. A novel prospective isolation of murine fetal liver progenitors to study in utero hematopoietic defects. PLoS Genet 14:e1007127 (2018). WB ; Mouse . Read more (PubMed: 29300724) »
  • Bergiers I  et al. Single-cell transcriptomics reveals a new dynamical function of transcription factors during embryonic hematopoiesis. Elife 7:N/A (2018). Read more (PubMed: 29555020) »
See all 27 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human glioblastoma cell line U87MG)
Specification
human glioblastoma cell line U87MG
Fixative
Paraformaldehyde
Permeabilization
Yes - 1% Triton X 100 in PBS
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 06 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (glioblastoma cell line from rat)
Specification
glioblastoma cell line from rat
Fixative
Paraformaldehyde
Permeabilization
Yes - 1% Triton X 100 in PBS
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 06 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (glioblastoma cell line C6)
Specification
glioblastoma cell line C6
Fixative
Paraformaldehyde
Permeabilization
Yes - 0,1% Triton X100 in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 24 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human glioblastoma cell line)
Specification
human glioblastoma cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0,1% Triton X100 in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 24 2012

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

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