Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21781] to NF-kB p65 (acetyl K310) - ChIP Grade
  • Suitable for: WB, Dot blot, ChIP
  • Reacts with: Human

Overview

  • Product name

    Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade
    See all NF-kB p65 primary antibodies

  • Description

    Rabbit monoclonal [EPR21781] to NF-kB p65 (acetyl K310) - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    WB
    Human
    Recombinant fragment
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HEK-293 transfected with NF-kB p65 and p300 (aa1287-1663) expression vectors containing a myc-His-tag®, then treated with 20 ng/ml TNF-alpha (ab9642) for 60 minutes whole cell lysate; Human lung tissue lysate; acetylated p65 purified protein. ChIP: Chromatin prepared from HeLa 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

  • Western blot - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)
    Western blot - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)
    Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533) at 1/1000 dilution + Human lung tissue lysate at 20 µg

    Secondary
    VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 60 kDa
    Observed band size: 65 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    We used a high sensitivity ECL substrate to develop this blot.

  • Western blot - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)
    Western blot - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533) This image is courtesy of an anonymous collaborator.
    Lanes 1-2 : Coomassie stain
    Lanes 3-4 : Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533) at 1/1000 dilution

    Lanes 1 & 3 : Acetylated p65 protein
    Lanes 2 & 4 : K310R mutant protein

    Lysates/proteins at 2 µg per lane.

    Secondary
    Lanes 3-4 : IRDye® 800CW Goat anti-Rabbit IgG (H + L) at 1/15000 dilution

    Predicted band size: 60 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?



    The p65 band runs higher in this blot because the protein contains a myc-tag.

  • ChIP - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)
    ChIP - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)

    Chromatin was prepared from HeLa (human epithelial cell line from cervix adenocarcinoma) cells treated with and without 20 ng/ml TNF-α (ab9642) for 60 minutes according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 μg of chromatin, 5 μg of ab218533 (red), and 20 µl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (SYBR green approach).

    The ChIP data are consistent with the literature (PMID: 16135789).

  • Western blot - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)
    Western blot - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)
    All lanes : Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533) at 1/2000 dilution

    Lane 1 : HEK-293 transfected with NF-kB p65 expression vector containing a myc-His-tag®, whole cell lysate
    Lane 2 : HEK-293 transfected with NF-kB p65 and p300 (aa1287-1663) expression vectors containing a myc-His-tag®, then treated with 20 ng/ml TNF-alpha (ab9642) for 60 minutes, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 60 kDa
    Observed band size: 70 kDa why is the actual band size different from the predicted?


    Exposure time: 37 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    NF-κB p65 (acetyl K310) expression is induced by TNF-α and p300 acetyltransferases (PMID: 20160011, PMID: 12456660, PMID: 16135789).

  • Dot Blot - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)
    Dot Blot - Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)

    Dot blot analysis of NF-kB p65 (acetyl K310) labeled with ab218533 at 1/1000 dilution.

    Lane 1: NF-κB p65 (acetyl K310) peptide.
    Lane 2: NF-κB p65 non-acetyl peptide.

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 2 minutes.

  • Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)
    Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)

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