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Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade (ab272456)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23309-113] to RUNX1 / AML1 - ChIP Grade
  • Suitable for: Flow Cyt, WB, ChIP, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade
    See all RUNX1 / AML1 primary antibodies

  • Description

    Rabbit monoclonal [EPR23309-113] to RUNX1 / AML1 - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Jurkat, MOLT-4 and THP-1 whole cell lysates. Flow Cyt: Jurkat cells. IP: Jurkat and K562 whole cell lysate.ChIP: Chromatin prepared from K562 cells.

  • General notes

    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.

Properties

Images

  • ChIP - Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade (ab272456)
    ChIP - Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade (ab272456)

    Chromatin was prepared from K-562 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab272456 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
    Primers and probes are from paper PMID：20959602

    *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

    The RUNX1 enrichment profile is consistent with what have been described in literature (PMID: 20959602).

  • Western blot - Anti-RUNX1 / AML1 antibody [EPR23309-113] (ab272456)
    Western blot - Anti-RUNX1 / AML1 antibody [EPR23309-113] (ab272456)
    All lanes : Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade (ab272456) at 1/1000 dilution

    Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : MOLT-4 (human lymphoblastic leukemia t lymphoblast) whole cell lysate
    Lane 3 : THP-1 (human monocytic leukemia monocyte) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 48 kDa
    Observed band size: 27-48 kDa
    why is the actual band size different from the predicted?



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

    RUNX1 has several isoforms. The molecular weight observed is consistent with what have been described in literature (PMID:17431130, 29296779). 

    Exposure time: 3 minutes

  • Immunoprecipitation - Anti-RUNX1 / AML1 antibody [EPR23309-113] (ab272456)
    Immunoprecipitation - Anti-RUNX1 / AML1 antibody [EPR23309-113] (ab272456)

    RUNX1 / AML1 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab272456 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab272456 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate 20 ug

    Lane 2: ab272456 IP in Jurkat whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab272456 in Jurkat whole cell lysate

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

    Exposure time: 32 seconds.

     

  • Flow Cytometry - Anti-RUNX1 / AML1 antibody [EPR23309-113] (ab272456)
    Flow Cytometry - Anti-RUNX1 / AML1 antibody [EPR23309-113] (ab272456)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) cells labelling RUNX1 / AML1 with ab272456 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade (ab272456)
    Immunoprecipitation - Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade (ab272456)

    RUNX1 / AML1 was immunoprecipitated from K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab272456 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272456 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: K562 whole cell lysate 10 µg

    Lane 2: ab272456 IP in K562 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab272456 in K562 whole cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 mins.

  • Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade (ab272456)
    Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade (ab272456)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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