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EpiTect ChIP OneDay Kit

For fast and effective chromatin immunoprecipitation

  • ChIP DNA from cells in just 6 hours
  • High-yield, high-quality DNA for accurate results
  • Compatible with any ChIP-grade antibody
  • For real-time PCR analysis and reproducible enrichment

The EpiTect ChIP OneDay Kit is designed for performing chromatin immunoprecipitation (ChIP) in 6 hours with a ChIP-grade antibody.

Cat No./ID: 334471
EpiTect ChIP OneDay Kit (12)
€280.00
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ChampionChIP One-Day Kit
The EpiTect ChIP OneDay Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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A high degree of reproducibility.

Four independent researchers collected duplicate samples of 2 million HCT116 cells. Each performed ChIP with RNA Polymerase II antibody (RNA Pol II) or control IgG from an EpiTect ChIP Antibody Kit. The specific enrichment of ChIP DNA was analyzed using an EpiTect ChIP qPCR Assay for the GAPDH proximal promoter. The results are expressed as percentages of input results (table), their average (chart y-axis), and standard deviation (error bars). Multiple researchers can achieve ChIP-qPCR of Pol II that agrees with only a 4.8% coefficient of variation.

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Good signal-to-noise ratios.

Triplicate wells of HCT-116 cells were fixed with formaldehyde. Chromatin was immunoprecipitated using the EpiTect ChIP OneDay Kit. Enriched genomic DNA was then purified and characterized by EpiTect ChIP qPCR Assays for the proximal promoter region of GAPDH and for an ORF-free region as a negative control. The results for each immunoprecipitation are expressed as percentage of input, or the fraction of the total input DNA co-immunoprecipitating with RNA Polymerase II.

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EpiTect ChIP OneDay protocol.
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DNA free from PCR inhibitors.
Chromatin was isolated from 4 million untreated HeLa cells and sonicated. DNA was then purified from an input DNA fraction (diamonds), and immunoprecipitations using either control IgG (triangles), anti-p53 (p53 IP, squares), or RNA Polymerase II (Pol2 IP, circles) using the EpiTect ChIP OneDay Kit. Different volumes of each DNA preparation (1.0, 2.0, and 4.0 µl) were used to obtain CT values and generate a calibration curve. Straight lines indicate that no inhibitors are present in the DNA preparations that interfere with real-time PCR. The results also demonstrate the reliability of the enrichment by the EpiTect ChIP OneDay Kit, since the fold of enrichment is independent of the volumes of template from the immunopreciptated DNA.
Performance
The EpiTect ChIP OneDay Kit isolates DNA with high yield and high purity, free from inhibitors. As a result, real-time PCR analysis of the DNA provides highly reproducible results and good signal-to-noise ratios. See figures "A high degree of reproducibility", "Good signal-to-noise ratios", and "DNA free from PCR inhibitors".
Principle

The EpiTect ChIP OneDay Kit is designed for performing chromatin immunoprecipitation (ChIP) using chosen antibodies to specifically enrich genomic DNA fragments bound in vivo to transcription factors, modified histones, or other nuclear factors of interest.

The EpiTect ChIP OneDay Kit decreases the time required for cross-link reversal and DNA isolation down to 1 hour versus traditional ChIP methods that require 6–8 hours. This time saving allows the rapid quality control and assessment of the size of sheared chromatin before immunoprecipitation, and enables more reliable subsequent real-time PCR detection.

It takes just 6 hours to generate a high yield of enriched ChIP DNA that is free of inhibitors and ready for real-time PCR quantification (see flowchart "EpiTect ChIP OneDay protocol"). The kit includes all the buffers and reagents needed for the ChIP preparative process, except for cross-linking agent and antibodies.

Procedure

Using the EpiTect ChIP OneDay Kit, chromatin from cells or tissues is first sheared and then incubated and immunoprecipitated with ChIP-grade antibodies from an EpiTect ChIP Antibody Kit or other sources. These antibodies enable precipitation of chromatin associated with the protein of interest. After purifying the ChIP DNA fraction with the EpiTect ChIP OneDay Kit, the amount of DNA enriched by the antibody of interest can be quantified by real-time PCR using EpiTect ChIP qPCR Assays or EpiTect ChIP qPCR Arrays.

Applications

Among other applications, the EpiTect ChIP OneDay Kit, used with the EpiTect ChIP Antibody Kit and EpiTect ChIP qPCR Arrays and Assays, streamlines:

Identification of disease biomarkers
Characterization of signal transduction pathways
Examination of transcriptional regulation
Detection of epigenetic changes
Verification of ChIP-on-chip and ChIP-Seq data

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Kit Handbooks (1)
For immunoprecipitation, crosslink reversal, and DNA purification for quantitative real-time PCR analysis of chromatin immunoprecipitation
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