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EpiQuik HAT Activity/Inhibition Assay Kit

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Suggested Workflow
Nuclear Protein Extraction
 
 
HAT Assay
 
Schematic procedure for using the EpiQuik HAT Activity/Inhibition Assay Kit.
Illustrated standard curve generated with HAT assay standard.
Nuclear proteins were prepared from different cell lines and HAT activity was measured using the EpiQuik™ HAT Activity/Inhibition Assay Kit.
Input Type: Nuclear Extracts, Purified Enzyme
Research Area: Histone Acetylation
Target Application: Activity Measurement
Vessel Format: 96-Well Plate
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-4003-4848 assays $221.00 
P-4003-9696 assays $399.00 
Availability: Usually Ships In 1-2 Days 
Product Overview

The EpiQuik™ HAT Activity/Inhibition Assay Kit is a ready-to-use set of essential components needed to measure histone acetyltransferase activity/inhibition safely and quickly. It is suitable for measuring histone acetyltransferase activity/inhibition from a broad range of species including mammalian cells/tissues, plants, and bacteria. The kit has the following advantages:

  • Very quick procedure, which can be completed within only 3 hours.
  • Innovative colorimetric assay without radioactivity, extraction, and chromatography.
  • Direct measurement of HAT activity and inhibition by quantifying the amount of acetylated histone substrate, thereby avoiding the false inhibitory effect on HATs.
  • Strip microplate format makes the assay flexible in both manual and high throughput analysis formats.
  • Simple, reliable, and consistent assay conditions.

Principle & Procedure
The EpiQuik™ HAT Activity/Inhibition Assay Kit is designed for measuring total histone aceyltransferase activity or inhibition. In an assay with this kit, the unique histone substrate is stably captured on the strip wells. Active HATs bind to and acetylate histone substrate. The acetylated substrate can be recognized with a high affinity anti-acetylated histone antibody. The ratio or amount of the acetylated histone, which is directly proportional to HAT enzyme activity, can then be colorimetrically quantified through an ELISA-like reaction.

 
Fig. 1. Schematic procedure for using the EpiQuik HAT Activity/Inhibition Assay Kit.

Fig. 2. Illustrated standard curve generated with HAT assay standard.

Fig. 3. Nuclear proteins were prepared from different cell lines and HAT activity was measured using the EpiQuik™ HAT Activity/Inhibition Assay Kit.

Product Components

HT1 (10X Wash Buffer)
HT2 (HAT Substrate, 20 µg/ml)*
HT3 (HAT Standard, 20 µg/ml)*
HT4 (Acetyl Co-A, 30 mM)*
HT5 (HAT Assay Buffer)
HT6 (Capture Antibody, 100 µg/ml)*
HT7 (Detection Antibody, 200 µg/ml)*
HT8 (Developing Solution)
HT9 (Stop Solution)
8-Well Assay Strip (with Frame)
User Guide

*For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

Product Citations

Jesse Roman et. al. (July 2017). Epigenetic regulation of EC-SOD expression in aging lung fibroblasts: role of histone acetylation Free Radical Biology and Medicine.

Roman J et. al. (July 2017). Epigenetic regulation of EC-SOD expression in aging lung fibroblasts: Role of histone acetylation. Free Radic Biol Med. 112:212-223.

Guan M et. al. (June 2017). Fatty acid synthase reprograms the epigenome in uterine leiomyosarcomas. PLoS One. 12(6):e0179692.

Večeřa J et. al. (March 2017). HDAC1 and HDAC3 Underlie Dynamic H3K9 Acetylation During Embryonic Neurogenesis and in Schizophrenia-like Animals. J Cell Physiol.

Ignácio ZM et. al. (March 2017). Quetiapine treatment reverses depressive-like behavior and reduces DNA methyltransferase activity induced by maternal deprivation. Behav Brain Res. 320:225-232.

Kuwabara T et. al. (October 2016). Acetylation Modulates IL-2 Receptor Signaling in T Cells. J Immunol.

Singh T et. al. (June 2016). Therapeutic intervention of silymarin on the migration of non-small cell lung cancer cells is associated with the axis of multiple molecular targets including class 1 HDACs, ZEB1 expression, and restoration of miR-203 and E-cadherin expression. Am J Cancer Res. 6(6):1287-301.

Nozik-Grayck E et. al. (May 2016). Histone deacetylation contributes to low extracellular superoxide dismutase expression in human idiopathic pulmonary arterial hypertension. Am J Physiol Lung Cell Mol Physiol. :ajplung.00263.2015.

Montagud-Romero S et. al. (May 2016). Up-regulation of histone acetylation induced by social defeat mediates the conditioned rewarding effects of cocaine. Prog Neuropsychopharmacol Biol Psychiatry.

Kala R et. al. (May 2016). A Novel Combinatorial Epigenetic Therapy Using Resveratrol and Pterostilbene for Restoring Estrogen Receptor-α (ERα) Expression in ERα-Negative Breast Cancer Cells. PLoS One. 11(5):e0155057.

Bhargavan B et. al. (November 2015). Toll-like receptor-3 mediates HIV-1 transactivation via NFκB and JNK pathways and histone acetylation, but prolonged activation suppresses Tat and HIV-1 replication. Cell Signal.

Chen Y et. al. (August 2015). Prenatal glucocorticoid contributed to rat lung dysplasia is related to asymmetric dimethylarginine/nitric oxide pathway Science Bulletin. 60(16):1416-1425.

Passacquale G et. al. (March 2015). Aspirin-induced histone acetylation in endothelial cells enhances synthesis of the secreted isoform of netrin-1 thus inhibiting monocyte vascular infiltration. Br J Pharmacol.

Prasad R et. al. (January 2015). Polyphenols from green tea inhibit the growth of melanoma cells through inhibition of class I histone deacetylases and induction of DNA damage. Genes Cancer. 6(1-2):49-61.

Chao MW et. al. (September 2014). Protective effects of ascorbic acid against the genetic and epigenetic alterations induced by 3,5-dimethylaminophenol in AA8 cells. J Appl Toxicol.

Ha SD et. al. (August 2014). HDAC8-Mediated Epigenetic Reprogramming Plays a Key Role in Resistance to Anthrax Lethal Toxin-Induced Pyroptosis in Macrophages. J Immunol. 193(3):1333-43.

Kumar P et. al. (June 2014). All-trans retinoic acid and sodium butyrate enhance natriuretic peptide receptor a gene transcription: role of histone modification. Mol Pharmacol. 85(6):946-57.

Siuda D et. al. (April 2014). Social isolation-induced epigenetic changes in midbrain of adult mice. J Physiol Pharmacol. 65(2):247-55.

Kumar P et. al. (March 2014). Histone deacetylase inhibitors modulate the transcriptional regulation of guanylyl cyclase/natriuretic peptide receptor-a gene: interactive roles of modified histones, histone acetyltransferase, p300, AND Sp1. J Biol Chem. 289(10):6991-7002.

Jenke A et. al. (January 2014). Restitution of gene expression and histone acetylation signatures altered by hepatitis B virus through antiviral microRNA-like molecules in nontransformed murine hepatocytes Clinical Epigenetics. 6:26. Full Article

Grissom NM et. al. (October 2013). Gestational overgrowth and undergrowth affect neurodevelopment: similarities and differences from behavior to epigenetics. Int J Dev Neurosci. 31(6):406-14.

Nayebosadri A et. al. (August 2013). Endothelial nuclear lamina is not required for glucocorticoid receptor nuclear import but does affect receptor-mediated transcription activation. Am J Physiol Cell Physiol. 305(3):C309-22.

Bhargavan B et. al. (March 2013). Epigenetic repression of LEDGF during UVB exposure by recruitment of SUV39H1 and HDAC1 to the Sp1-responsive elements within LEDGF promoter CpG island. Epigenetics. 8(3):268-80.

Singh T et. al. (January 2013). Inhibition of class I histone deacetylases in non-small cell lung cancer by honokiol leads to suppression of cancer cell growth and induction of cell death in vitro and in vivo. Epigenetics. 8(1):54-65.

Xiong L et. al. (December 2012). DNA demethylation regulates the expression of miR-210 in neural progenitor cells subjected to hypoxia. FEBS J. 279(23):4318-26.

Pascual M et. al. (June 2012). Changes in histone acetylation in the prefrontal cortex of ethanol-exposed adolescent rats are associated with ethanol-induced place conditioning. Neuropharmacology. 62(7):2309-19.

Meeran SM et. al. (May 2012). Bioactive dietary supplements reactivate ER expression in ER-negative breast cancer cells by active chromatin modifications. PLoS One. 7(5):e37748.

Nishino T et. al. (September 2011). Ex vivo expansion of human hematopoietic stem cells by garcinol, a potent inhibitor of histone acetyltransferase. PLoS One. 6(9):e24298.

Kraft M et. al. (September 2011). Disruption of the histone acetyltransferase MYST4 leads to a Noonan syndrome-like phenotype and hyperactivated MAPK signaling in humans and mice. J Clin Invest. 121(9):3479-91.

Meeran SM et. al. (August 2011). A novel prodrug of epigallocatechin-3-gallate: differential epigenetic hTERT repression in human breast cancer cells. Cancer Prev Res (Phila). 4(8):1243-54.

Czakai K et. al. (August 2011). Perturbation of mitosis through inhibition of histone acetyltransferases: the key to ochratoxin a toxicity and carcinogenicity? Toxicol Sci. 122(2):317-29.

Pascual M et. al. (June 2011). Impact of TLR4 on behavioral and cognitive dysfunctions associated with alcohol-induced neuroinflammatory damage. Brain Behav Immun. 25 Suppl 1:S80-91.

Sui L et. al. (December 2010). Effects of perinatal hypothyroidism on regulation of reelin and brain-derived neurotrophic factor gene expression in rat hippocampus: Role of DNA methylation and histone acetylation. Steroids. 75(12):988-97.

Li Y et. al. (October 2010). Synergistic epigenetic reactivation of estrogen receptor-α (ERα) by combined green tea polyphenol and histone deacetylase inhibitor in ERα-negative breast cancer cells. Mol Cancer. 9:274.

Meeran SM et. al. (July 2010). Sulforaphane causes epigenetic repression of hTERT expression in human breast cancer cell lines. PLoS One. 5(7):e11457.

Majid S et. al. (January 2010). Genistein reverses hypermethylation and induces active histone modifications in tumor suppressor gene B-Cell translocation gene 3 in prostate cancer. Cancer. 116(1):66-76.

Watson JA et. al. (October 2009). Generation of an epigenetic signature by chronic hypoxia in prostate cells. Hum Mol Genet. 18(19):3594-604.

Majid S et. al. (April 2009). BTG3 tumor suppressor gene promoter demethylation, histone modification and cell cycle arrest by genistein in renal cancer. Carcinogenesis. 30(4):662-70.

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