Overview

  • Product name
    Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric)
  • Detection method
    Fluorescent
  • Sample type
    Cell culture extracts, Tissue Extracts
  • Assay type
    Enzyme activity
  • Assay time
    1h 00m
  • Species reactivity
    Reacts with: Human
  • Product overview

    Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) ab156064 detects HDAC activity in lysates. Primarily, the Histone Deacetylase (HDAC) Activity Assay Kit is designed for the rapid and sensitive evaluation of HDAC inhibitors using crude HDAC fraction. Additionally, any cultured primary cell, cell line, or tissue homogenate can be assayed for HDAC activity with the Histone Deacetylase (HDAC) Activity Assay Kit if the appropriate dose of HDAC specific inhibitor e.g. Trichostatin A is used.


    The Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) has been shown to detect the activity of HDAC family, at least class I HDACs in Human or animal cell lysates or in column fractions. The assay shows good linearity of sample response. The assay may be used to follow the purification of HDACs or may be used to detect the presence of HDACs in cell lysates


    Applications for this kit include:


    1. Monitoring the purification of HDACs including HDAC1, 2, 3 and 8.


    2. Screening inhibitors or activators of HDACs.


    3. Detecting the effects of pharmacological agents on HDACs.

  • Notes

    Histone Deacetylases (HDACs) are a class of enzymes responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4), allowing the histones to wrap the DNA more tightly.

    HDAC proteins occur in four groups (class I, class IIA, class IIB, class III, class IV) based on function and DNA sequence similarity.
    Classes I, IIA and IIB are considered "classical" HDACs whose activities are inhibited by trichostatin A (TSA), whereas class III is a family of NAD+-dependent proteins (sirtuins) not affected by TSA. Class IV is considered an atypical class on its own, based solely on DNA sequence similarity to the others.

     

Properties

  • Storage instructions
    Please refer to protocols.
  • Components 100 tests
    Crude HDAC 1 x 500µl
    Developer 1 x 500µl
    Fluoro-Deacetylated Peptide (0.2 mM) 1 x 100µl
    Fluoro-Substrate Peptide (0.2 mM) 1 x 500µl
    HDAC Assay Buffer 2 x 1ml
    HDAC Stop Solution 2 x 1ml
    Trichostatin A (10µM) 1 x 500µl
  • Research areas
  • Function
    Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Deacetylates SP proteins, SP1 and SP3, and regulates their function. Component of the BRG1-RB1-HDAC1 complex, which negatively regulates the CREST-mediated transcription in resting neurons. Upon calcium stimulation, HDAC1 is released from the complex and CREBBP is recruited, which facilitates transcriptional activation. Deacetylates TSHZ3 and regulates its transcriptional repressor activity. Deacetylates 'Lys-310' in RELA and thereby inhibits the transcriptional activity of NF-kappa-B. Component a RCOR/GFI/KDM1A/HDAC complex that suppresses, via histone deacetylase (HDAC) recruitment, a number of genes implicated in multilineage blood cell development.
  • Tissue specificity
    Ubiquitous, with higher levels in heart, pancreas and testis, and lower levels in kidney and brain.
  • Sequence similarities
    Belongs to the histone deacetylase family. HD type 1 subfamily.
  • Post-translational
    modifications
    Sumoylated on Lys-444 and Lys-476; which promotes enzymatic activity. Desumoylated by SENP1.
    Phosphorylation on Ser-421 and Ser-423 promotes enzymatic activity and interactions with NuRD and SIN3 complexes. Phosphorylated by CDK5.
    Ubiquitinated by CHFR, leading to its degradation by the proteasome (By similarity). Ubiquitinated by KCTD11, leading to proteasomal degradation.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Alternative names
    • HD1
    • HDAC1
    • HDAC1_HUMAN
    • Histone deacetylase 1
  • Database links

Images

  • 8 uL (corresponding to 6.7e6 cells) of nuclear extracts (ab113474) from HL60 cells were assessed kinetically (+/- SD).

  • Different volumes of positive control, with inhibitor (TA), kinetically read (+/- SD).

  • Measurement of HeLa cell endogenous HDAC1 in an immunoprecipitated sample with anti-HDAC1 antibody and following the Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) (ab156064) protocol.

  • Measurement of HeLa cell endogenous HDAC2 in an immunoprecipitated sample with anti-HDAC2 antibody following the Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) (ab156064) protocol.

Protocols

References

This product has been referenced in:
  • Alagarswamy K  et al. Region-specific alteration of histone modification by LSD1 inhibitor conjugated with pyrrole-imidazole polyamide. Oncotarget 9:29316-29335 (2018). Read more (PubMed: 30034620) »
  • Li M  et al. The Anti-inflammatory Effects of Short Chain Fatty Acids on Lipopolysaccharide- or Tumor Necrosis Factor a-Stimulated Endothelial Cells via Activation of GPR41/43 and Inhibition of HDACs. Front Pharmacol 9:533 (2018). Read more (PubMed: 29875665) »
See all 10 Publications for this product

Customer reviews and Q&As

Answer

Unfortunately, I cannot tell you which of HDACs can be measured with certainty, because the lab has measured ab156064 only with crude HDACs fractions of some cell lysates, and recombinant HDAC1, 2, 3 and 8.

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