KAT2B/pCAF Inhibitor Screening Assay Kit (Fluorometric) (ab196991)

Overview

  • Product name

    KAT2B/pCAF Inhibitor Screening Assay Kit (Fluorometric)
  • Detection method

    Fluorescent
  • Sample type

    Inhibitor compounds
  • Assay type

    Enzyme activity
  • Product overview

    KAT2B/pCAF Inhibitor Screening Assay Kit (Fluorometric) (ab196991) is an assay where histone H3 peptide and Acetyl CoA are used as acetylation substrates. KAT2B/pCAF acetylates the Histone H3 peptide and generates Coenzyme A with a free thiol group (CoA-SH). CoA-SH  reacts subsequently with a Thiol Detecting Probe in order to increase the amount of fluorescence in the solution, which can be measured at Ex/Em = 392/482 nm. In the presence of KAT2B/pCAF specific inhibitors, the enzymatic activity is reduced ot completely abolished resulting in decreased or total loss of fluorescence.

    This assay kit is a simple, sensitive and rapid tool to screen potential pCAF inhibitors.

  • Notes

    KTA2B (K-(lysine) acetyltransferase 2B, EC 2.3.1.48), also known as P300/CBP-associated factor (pCAF), possesses intrinsic histone acetyltransferase (HAT) activity and is able to acetylated both histone H3 and histone H4. KTA2B/pCAF contributes to transcriptional activation by acetylating chromatin and transcription factors through its HAT activity. Deregulated HAT activity plays an important role in various cancers.

    KTA2B/pCAF inhibitors have potential applications in cancer therapy.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Acetyl CoA 1 vial
    H3 Peptide 1 vial
    pCAF Assay Buffer 1 x 25ml
    pCAF Enzyme 1 x 40µl
    pCAF Inhibitor 1 x 10µl
    Thiol Detecting Probe 1 x 200µl
  • Function

    Functions as a histone acetyltransferase (HAT) to promote transcriptional activation. Has significant histone acetyltransferase activity with core histones (H3 and H4), and also with nucleosome core particles. Also acetylates non-histone proteins, such as ACLY. Inhibits cell-cycle progression and counteracts the mitogenic activity of the adenoviral oncoprotein E1A. 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.
  • Tissue specificity

    Ubiquitously expressed but most abundant in heart and skeletal muscle.
  • Sequence similarities

    Belongs to the acetyltransferase family. GCN5 subfamily.
    Contains 1 bromo domain.
    Contains 1 N-acetyltransferase domain.
  • Domain

    The bromodomain mediates binding to HIV-1 Tat.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Alternative names

    • Histone acetylase PCAF
    • Histone acetyltransferase KAT2B
    • Histone acetyltransferase PCAF
    • KAT2B
    • KAT2B_HUMAN
    • Lysine acetyltransferase 2B
    • P/CAF
    • P300/CBP-associated factor
    see all

Images

  • Inhibition of KAT2B/pCAF activity by different concentrations of the KAT2B/pCAF Inhibitor (Garcinol).

Protocols

References

ab196991 has not yet been referenced specifically in any publications.

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