Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876)

Overview

  • Product name

    Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade
    See all KAT2A / GCN5 primary antibodies
  • Description

    Rabbit monoclonal [EPR21146] to KAT2A / GCN5 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ChIP, WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human KAT2A/ GCN5 aa 50-350. The exact sequence is proprietary.
    Database link: Q92830

  • Positive control

    • WB: HAP1, HEK-293T, MCF7 and HeLa whole cell lysate; human fetal brain tissue lysate; His-tagged human KAT2A / GCN5 recombinant protein (aa86-336). IP: HeLa whole 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 ab217876 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.
WB 1/1000. Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa).
IP 1/30.

Target

  • Function

    Functions as a histone acetyltransferase (HAT) to promote transcriptional activation. Acetylation of histones gives a specific tag for epigenetic transcription activation. Has significant histone acetyltransferase activity with core histones, but not with nucleosome core particles. In case of HIV-1 infection, it is recruited by the viral protein Tat. Regulates Tat's transactivating activity and may help inducing chromatin remodeling of proviral genes. Component of the ATAC complex, a complex with histone acetyltransferase activity on histones H3 and H4.
  • Tissue specificity

    Expressed in all tissues tested, with most abundant expression in ovary.
  • Sequence similarities

    Belongs to the GCN5 family.
    Contains 1 bromo domain.
    Contains 1 N-acetyltransferase domain.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • 1110051E14Rik antibody
    • AW212720 antibody
    • EC 2.3.1.48 antibody
    • GCN 5 antibody
    • GCN5 (general control of amino-acid synthesis, yeast, homolog)-like 2 antibody
    • Gcn5 antibody
    • GCN5 general control of amino-acid synthesis 5-like 2 (yeast) antibody
    • GCN5L2 antibody
    • general control of amino acid synthesis 5-like 2 antibody
    • General control of amino acid synthesis protein 5-like 2 antibody
    • General control of amino acid synthesis, yeast, homolog-like 2 antibody
    • HGCN5 antibody
    • Histone acetyltransferase GCN5 antibody
    • Histone acetyltransferase KAT2A antibody
    • hsGCN5 antibody
    • K(lysine) acetyltransferase 2A antibody
    • KAT2 A antibody
    • KAT2A antibody
    • KAT2A_HUMAN antibody
    • Lysine acetyltransferase 2A antibody
    • MGC102791 antibody
    • PCAF-b antibody
    • STAF97 antibody
    see all

Images

  • All lanes : Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876) at 1/1000 dilution

    Lane 1 : His-tagged human KAT2A / GCN5 recombinant protein (aa86-336), 10 ng
    Lane 2 : His-tagged human KAT2B recombinant protein (aa74-326), 10 ng

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

    Developed using the ECL technique.

    Predicted band size: 94 kDa
    Observed band size: 27 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST

    Exposure time: 1 second

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

  • Chromatin was prepared from A549 treated with thapsigargin (1µM 12 hours) cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab217876 (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).
    Primers and probes are located in the first kb of the transcribed region.
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
  • KAT2A / GCN5 was immunoprecipitated from 0.35mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab217876 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217876 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: HeLa whole cell lysate 10ug (Input). 

    Lane 2: ab217876 IP in HeLa whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab217876 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 30 seconds.

  • All lanes : Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876) at 1/1000 dilution

    Lane 1 : Wild type HAP1 whole cell lysate
    Lane 2 : KAT2A / GCN5-knockout HAP1 whole cell lysate whole cell lysate
    Lane 3 : HEK-293T (uman epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 94 kDa
    Observed band size: 94 kDa


    Exposure time: 3 minutes


    ab217876 was shown to specifically react with KAT2A / GCN5 in wild-type HAP1 cells as signal was lost in KAT2A / GCN5 knockout cells. Wild-type and KAT2A / GCN5 knockout samples were subjected to SDS-PAGE. Ab217876 and ab181602 (Human anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/200000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrumentusing the ECL technique.

  • All lanes : Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Human fetal brain tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lane 2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 94 kDa
    Observed band size: 94 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST

    Exposure time: Lane 1: 15 seconds; Lane 2: 3 minutes

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

References

ab217876 has not yet been referenced specifically in any publications.

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