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EpiQuik Chromatin Accessibility Assay Kit

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For quantitative chromatin analysis and determination of euchromatin or heterochromatin states

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Schematic procedure of the EpiQuik™ Chromatin Accessibility Assay Kit.
Chromatin accessibility of opened chromatin (euchromatin). Amplification of proximal promoter regions for the constitutively expressed target gene was carried out in MDA-231 cells by using positive control primers and using the EpiQuik™ Chromatin Accessibility Assay Kit (Cat. #P-1047) and EpiQuik™ Quantitative PCR Fast Kit (Cat. #P-1029). Red and green lines: Nse-untreated; Blue lines: Nse-treated. The difference of Ct values between Nse-treated and untreated is > 7 cycles.
Chromatin accessibility of closed chromatin (heterochromatin). Amplification of proximal promoter regions for the constitutively repressed target gene was carried out in MDA-231 cells by using negative control primers and using the EpiQuik™ Chromatin Accessibility Assay Kit (Cat #P-1047) and EpiQuik™ Quantitative PCR Fast Kit (Cat #P-1029). Red and green lines: Nse-untreated; Blue and pink lines: Nse-treated. The difference of Ct values between Nse-treated and untreated is <1.5 cycles.
Input Type: Cell Extracts, Chromatin, Unisolated Samples
Research Area: Chromatin & Transcription
Target Application: Sample Modification
Vessel Format: Columns/Tubes
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-1047-4848 assays $489.00 
Order within 10 hr 39 min & get it by tomorrow 
Product Overview

The EpiQuik™ Chromatin Accessibility Assay Kit is a complete set of optimized reagents designed for conducting a gene-specific analysis of chromatin accessibility including nucleosome/transcription factor positioning from various biological samples via real time PCR. It is a cost-effective and useful tool for quick detection of open and closed chromatin. The kit has the following advantages and features:

  • Extremely fast and convenient protocol that allows the entire procedure (from cell tissue sample to ready-to-use DNA for PCR) to be finished in as short as 1 hour and 30 minutes.
  • Able to be used with cultured cells and also fresh and frozen tissues.
  • Fast process minimizes nuclear damage and loss of disassociated chromatin components, preserving chromatin structure.
  • Internal control primers for human are included in the kit as references for analyzing the extent of chromatin accessibility in a specified gene target in the sample DNA, and also for validating whether the proper enzymatic digestions are achieved.
  • Choice of single-reaction assay or high throughput multi-reaction assay can be used, making the assay flexible.

Background Information
The accessibility of regulatory elements in chromatin is critical for many aspects of gene regulation. Nucleosomes positioned over regulatory elements inhibit access of transcription factors to DNA. To elucidate the role of the interactions between chromatin and transcription factors, it is crucial to determine chromatin accessibility through mapping of the nucleosome positioning along the genome. In general, the more condensed the chromatin, the more difficult it is for transcription factors and other DNA binding proteins to access DNA and carry out their tasks. The more accessible the DNA, the more likely surrounding genes are actively transcribed. The presence (or the absence) of nucleosomes directly or indirectly affects a variety of other cellular and metabolic processes such as recombination, replication, centromere formation, and DNA repair.

Principle & Procedure
The EpiQuik™ Chromatin Accessibility Assay Kit contains all the necessary reagents required for obtaining a gene-specific analysis of chromatin accessibility from cell/tissue samples via real time PCR. This kit includes internal control primers for determining whether chromatin digestion is successfully achieved. In this assay, chromatin is isolated from the cells/tissues and is treated with a nuclease mix. DNA is then isolated and amplified with real time PCR for region-specific analysis of chromatin accessibility. Chromatin states can be identified based on how accessible the DNA is to nucleases. The DNA in heterochromatin is inaccessible to outside proteins, including exogenous nucleases, rendering it protected by nucleosome or DNA/protein complexes and becomes available for subsequent PCR with insignificant Ct shifts between digested and undigested samples. In contrast, the DNA in euchromatin (nucleosome depletion) is accessible to exogenous nucleases, making it susceptible to nuclease digestion and becomes unavailable for PCR with a large Ct shift between digested and undigested samples. 

Starting Materials & Input Amount
Starting materials can include various mammalian tissue or cell samples such as cells from flask or microplate cultured cells, fresh and frozen tissues, etc. The amount of cell/tissues for each reaction can be from 1 x 105 cells or 2 mg tissues to 2 x 106 cells or 40 mg tissues. For an optimal reaction, the input chromatin amount should be about 0.5 x 106 cells or 10 mg tissues.

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.

[Safety Data Sheet]
Product Citations

Khanal T et. al. (April 2019). NR2E3 is a key component in p53 activation by regulating a long noncoding RNA DINO in acute liver injuries. FASEB J. :fj201801881RR.

Khanal T et. al. (April 2019). NR2E3 is a key component in p53 activation by regulating a long noncoding RNA DINO in acute liver injuries. FASEB J. :fj201801881RR.

Urdinguio RG et. al. (March 2019). Chromatin regulation by Histone H4 acetylation at Lysine 16 during cell death and differentiation in the myeloid compartment. Nucleic Acids Res.

Li M et. al. (January 2019). Mono-ADP-ribosylation of H3R117 traps 5mC hydroxylase TET1 to impair demethylation of tumor suppressor gene TFPI2. Oncogene.

Sturrock A et. al. (December 2018). Consequences of Hypoxia for the Pulmonary Alveolar Epithelial Cell Innate Immune Response. J Immunol. 201(11):3411-3420.

Kim GC et. al. (November 2018). Upregulation of Ets1 expression by NFATc2 and NFKB1/RELA promotes breast cancer cell invasiveness. Oncogenesis. 7(11):91.

Koh B et. al. (November 2018). A conserved enhancer regulates Il9 expression in multiple lineages. Nat Commun. 9(1):4803.

Thakur C et. al. (September 2018). Loss of mdig expression enhances DNA and histone methylation and metastasis of aggressive breast cancer. Signal Transduct Target Ther. 3:25.

Noutsou M et. al. (September 2017). The Cohesin Complex Is Necessary for Epidermal Progenitor Cell Function through Maintenance of Self-Renewal Genes. Cell Rep. 20(13):3005-3013.

Jeon S et. al. (July 2017). Shift of EMT gradient in 3D spheroid MSCs for activation of mesenchymal niche function. Sci Rep. 7(1):6859.

Kinner-Bibeau LB et. al. (May 2017). HSPs drive dichotomous T-cell immune responses via DNA methylome remodelling in antigen presenting cells. Nat Commun. 8:15648.

Swathy B et. al. (April 2017). Pharmacoepigenomic responses of antipsychotic drugs on pharmacogenes are likely to be modulated by miRNAs. Epigenomics.

Krause BJ et. al. (November 2015). Arginase-2 is cooperatively up-regulated by nitric oxide and histone deacetylase inhibition in human umbilical artery endothelial cells. Biochem Pharmacol.

Vincent ZL et. al. (October 2015). Regulation of MT1-MMP/MMP-2/TIMP-2 axis in human placenta. J Inflamm Res. 8:193-200.

Customer Reviews

Rating by f*********@gmail.com Verified Purchase Reviewed on: Monday 13 May, 2019
Application Description
Assay chromatin accessibility of specific gene. As well as comparing the chromatin accessibility of two enzymes that made in house.

Pros Details
The protocol is straight forward and easy to follow. The procedures are fast and less labor intensity compared to other methods. The price is also cheaper compare to another vendor.

Cons Details
Lack some details and description in the procedures writing.

Observed white precipitate while eluting DNA from Column.
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