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EpiQuik HDAC Activity/Inhibition Assay Kit (Colorimetric)

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Schematic procedure for the EpiQuik HDAC Activity/Inhibition Assay Kit (Colorimetric).
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-4002-4848 assays $194.00 
P-4002-9696 assays $338.00 
Availability: Usually Ships In 1-2 Days 
Product Overview





We recommend using a newer version of this product: Epigenase HDAC Activity/Inhibition Direct Assay Kit (Colorimetric)




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

WHY CHOOSE THE EPIQUIK™ HDAC ACTIVITY/INHIBITION ASSAY KIT (COLORIMETRIC),

  • Very quick procedure, which can be completed within 3 hours.
  • Innovative colorimetric assay without radioactivity, extraction, and chromatography.
  • Direct measurement of HDAC activity and inhibition without the use of lysyl endopeptidase, thereby avoiding the false inhibitory effect on HDACs.
  • Strip microplate format makes the assay flexible in both manual and high throughput analysis formats.
  • Simple, reliable, and consistent assay conditions.
Product Details

The EpiQuik™ HDAC Activity/Inhibition Assay Kit is designed for measuring total HDAC activity/inhibition. In an assay with this kit, the unique acetylated histone substrate is stably captured on the strip wells. Active HDACs bind to and deacetylate histone substrate. The remaining un-deacetylated substrate can be recognized with a high affinity acetylated histone antibody. The ratio or amount of the un-deacetylated histone, which is inversely proportional to HDAC enzyme activity, can then be colorimetrically quantified through an ELISA-like reaction.

SCHEMATIC PROCEDURE:  



Product Components

H1 (10X Wash Buffer)
H2 (HDAC Assay Buffer)
H3 (Biotinylated HDAC Substrate)*
H4 (HDAC Inhibitor, 0.5 mM)*
H5 (HDAC Standard, 20 µg/ml)*
H6 (Capture Antibody, 100 µg/ml)*
H7 (Detection Antibody, 200 µg/ml)*
H8 (Developing Solution)
H9 (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.

Frequently Asked Q's

1. In what kind of medium should the nuclear extract be made?
Any nuclear extraction preparation methods can be used.

2. What classes of HDACs can your HDAC Activity/Inhibition Assay Kit (Cat. No. P-4002) be used for?
At least class 1 and 2 can be assayed with this kit.

3. How many cells should be used per test?
50,000-100,000 cells should be sufficient for each assay point.

4. When doing preparation of the standard curve, "add 50 µl of 1XH1 into the well, followed by adding 1 µl of H5 at a different amount (0.1-10 ng)." It had been mentioned adding 1 µl of H5, what does it mean by different amount (0.1-10 ng)?
Volume is 1 µl. The concentration should be different (several concentration points, ex: 0.1, 0.5, 2, 5, and 10 ng) to generate the dose-response standard curve.

5. In the formula of calculate HDAC activity, does the hour specifically refer to the incubating time 30-60min in step 5?
Yes, the hour here is the incubation time in step 5 (0.5-1 hour).

6. The main HDAC which I am going to detection is HDAC6.The substrate of HDAC6 is tubulin and HSP90. Can I use kit P-4002 to detect this?
Kit P-4002 would be suitable for HDAC6, as the substrate. Included in this kit is universal for class I and II HDACs.

7. In the protocol, the first step is to prepare nuclear extracts. In my experiment, the HDAC which I am going to detect is in the cytoplasm. Can I just use the cytoplasm for detecting HDAC?
You can use the cytoplasm for the HDAC detection, as some of HDACs are in both cytoplasm and nucleus, especially class II.

8. These kits are described as having specificity to a broad range of species. Can you tell me if these antibodies have been validated in any species in particular?
These antibodies have been validated in human, mouse, and rat. These antibodies are also expected to react with a broad range of species due to protein sequence homology.

9. Previously you mentioned that the reaction fluid can be transferred to cuvettes for colorimetric detection. This means that the detected solution is not fixed to the bottom of the plate. But how can you do the repeated wash steps, in this case?
The enzymatically converted product still stick on the strip well and can be repeatedly washed. However, it will generate the colorimetric signal through an ELISA-like reaction at the final step and cause the reaction solution to be colorful.

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.

[Material Safety Data Sheet]
Product Citations

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.

Nettersheim D et. al. (August 2016). A signaling cascade including ARID1A, GADD45B and DUSP1 induces apoptosis and affects the cell cycle of germ cell cancers after romidepsin treatment. Oncotarget.

Nettersheim D et. al. (August 2016). A signaling cascade including ARID1A, GADD45B and DUSP1 induces apoptosis and affects the cell cycle of germ cell cancers after romidepsin treatment. Oncotarget.

Li Y et. al. (August 2016). Combinatorial epigenetic mechanisms and efficacy of early breast cancer inhibition by nutritive botanicals. Epigenomics. 8(8):1019-37.

Eisses JF et. al. (October 2015). Valproic Acid Limits Pancreatic Recovery after Pancreatitis by Inhibiting Histone Deacetylases and Preventing Acinar Redifferentiation Programs. Am J Pathol.

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.

Borutinskaitė V et. al. (August 2015). The Histone Deacetylase Inhibitor BML-210 Influences Gene and Protein Expression in Human Promyelocytic Leukemia NB4 Cells via Epigenetic Reprogramming. Int J Mol Sci. 16(8):18252-69.

Berkholz J et. al. (July 2015). skNAC and Smyd1 in transcriptional control. Exp Cell Res.

Varghese TA et. al. (April 2015). Marine actinomycetes as potential source for histone deacetylase inhibitors and epigenetic modulation. Lett Appl Microbiol.

Zhu Y et. al. (February 2015). Alteration of Histone Acetylation Pattern during Long-Term Serum-Free Culture Conditions of Human Fetal Placental Mesenchymal Stem Cells. PLoS One. 10(2):e0117068.

Kim JH et. al. (September 2014). Histone acetylation level and histone acetyltransferase/deacetylase activity in ejaculated sperm from normozoospermic men. Yonsei Med J. 55(5):1333-40.

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.

Kirpich I et. al. (November 2013). Binge ethanol-induced HDAC3 down-regulates Cpt1α expression leading to hepatic steatosis and injury. Alcohol Clin Exp Res. 37(11):1920-9.

Toussirot E et. al. (August 2013). Imbalance between HAT and HDAC activities in the PBMCs of patients with ankylosing spondylitis or rheumatoid arthritis and influence of HDAC inhibitors on TNF alpha production. PLoS One. 8(8):e70939.

Yaacob JS et. al. (June 2013). Protein profiling and histone deacetylation activities in somaclonal variants of oil palm (Elaeis guineensis Jacq.). ScientificWorldJournal. 2013:613635.

Karaca B et. al. (June 2013). Combination of AT-101/cisplatin overcomes chemoresistance by inducing apoptosis and modulating epigenetics in human ovarian cancer cells. Mol Biol Rep. 40(6):3925-33.

Shah RD et. al. (April 2013). Sodium valproate potentiates staurosporine-induced apoptosis in neuroblastoma cells via Akt/survivin independently of HDAC inhibition. J Cell Biochem. 114(4):854-63.

Kauntz H et. al. (April 2013). Epigenetic effects of the natural flavonolignan silibinin on colon adenocarcinoma cells and their derived metastatic cells. Oncol Lett. 5(4):1273-1277.

Barcellos KS et. al. (January 2013). ARHGAP21 protein, a new partner of α-tubulin involved in cell-cell adhesion formation and essential for epithelial-mesenchymal transition. J Biol Chem. 288(4):2179-89.

Jeong JB et. al. (January 2013). Protocatechualdehyde possesses anti-cancer activity through downregulating cyclin D1 and HDAC2 in human colorectal cancer cells. Biochem Biophys Res Commun. 430(1):381-6.

Bozkurt E et. al. (November 2012). Effects of Thymus serpyllum extract on cell proliferation, apoptosis and epigenetic events in human breast cancer cells. Nutr Cancer. 64(8):1245-50.

Ling X et. al. (September 2012). A novel small molecule FL118 that selectively inhibits survivin, Mcl-1, XIAP and cIAP2 in a p53-independent manner, shows superior antitumor activity. PLoS One. 7(9):e45571.

Kirpich I et. al. (September 2012). Binge alcohol-induced microvesicular liver steatosis and injury are associated with down-regulation of hepatic Hdac 1, 7, 9, 10, 11 and up-regulation of Hdac 3. Alcohol Clin Exp Res. 36(9):1578-86.

Lanzillotta A et. al. (August 2012). Targeted acetylation of NF-kappaB/RelA and histones by epigenetic drugs reduces post-ischemic brain injury in mice with an extended therapeutic window. Neurobiol Dis. 49C:177-189.

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.

Trisciuoglio D et. al. (January 2012). CPTH6, a thiazole derivative, induces histone hypoacetylation and apoptosis in human leukemia cells. Clin Cancer Res. 18(2):475-86.

Shu L et. al. (December 2011). Epigenetic CpG demethylation of the promoter and reactivation of the expression of Neurog1 by curcumin in prostate LNCaP cells. AAPS J. 13(4):606-14.

Whitworth KM et. al. (June 2011). Scriptaid corrects gene expression of a few aberrantly reprogrammed transcripts in nuclear transfer pig blastocyst stage embryos. Cell Reprogram. 13(3):191-204.

Grabiec AM et. al. (March 2010). Histone deacetylase inhibitors suppress inflammatory activation of rheumatoid arthritis patient synovial macrophages and tissue. J Immunol. 184(5):2718-28.

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.

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.

Customer Reviews

Rating By L**@uab.edu Verified Purchase Reviewed on: Monday 19 December, 2016
Application Description
We used EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (p-3009) and EpiQuik HDAC Activity/Inhibition Assay Kit (p-4002) to check DNMT and HDAC activity in tumor cells.

Pros: They are very easy to work with. Protocols are clearly delineated the detailed procedures.
Cons: Too much steps and solutions, which may cause confusion and mistakes.

Other Thoughts
1) DNMT kit should have positive control included.
2) There is a concern for how to determine internal HDAC and DNMT quantity (protein level) to normalize all samples

How friendly was the user guide? (1 – worst, 5 – best)
4

How professional was the appearance and presentation of the product? (1 – worst, 5 – best)
5

How would you rate the overall product? (1 – worst, 5 – best)
4.5
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