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EpiTect Chip One-Day Kit (12)
Fast ChIP with Reliable Results

 
EpiTect Chip One-Day Kit (12)
Catalog No. Contents Price
334471 For 12 Chromatin Immunoprecipitations & DNA Purifications
The EpiTect Chip One-Day Kit is designed for performing chromatin immunoprecipitation (ChIP) using your carefully chosen antibodies to specifically enrich genomic DNA fragments that bind in vivo to transcription factors, modified histones or other nuclear factors of interest. The EpiTect Chip One-Day Kit decreases the time needed for cross-link reversal and DNA isolation down to one hour versus traditional ChIP methods that require six to eight hours. This innovation allows you to quickly assess the size of your sheared chromatin before immunoprecipitation, and to effectively recover enriched genomic DNA after immunoprecipitation. It therefore only takes 6 hours to generate a high yield of enriched ChIP DNA that is free of inhibitors and ready for real-time PCR quantification. The kit includes all the buffers and reagents needed for the ChIP preparative process except cross-linking agent and antibodies.

What the EpiTect Chip One-Day Kit Provides:

Speed & Ease: Simplified protocol easily obtains ChIP DNA from cells in just 6 hours
Universality: General kit compatible with any ChIP-Grade antibody
Reliability: High-yield and high-quality DNA for real-time PCR and highly reproducible enrichment

User Manual

 

How It Works Performance Related Products Materials Included
 
How It Works: Speed and Ease

The Entire EpiTect Chip System Protocol Can Be Completed in A Single Day. The EpiTect Chip System includes a simplified high-performance One-Day ChIP Kit, ChIP-Grade Antibodies, real-time PCR primers, and a FREE Excel-based data analysis template. The figure depicts how all the components of the EpiTect Chip System work together. The EpiTect Chip One-Day Kit simplifies the usual two- to five-day ChIP protocol down to six or eight hours. Its crosslink reversal step is much faster and less tedious than conventional methods, and its DNA purification step yields more ChIP DNA of higher quality than other one-day kits. These innovations help you quickly assess or validate the size of your sheared chromatin and efficiently recover DNA after every ChIP experiment. The genome-wide experimentally-validated real-time PCR primers now allows you to quantitatively analyze genomic DNA enrichment in ChIP samples with a level accuracy and throughput previously not possible with traditional agarose gel based analysis.

 

How It Works Performance (QC) Related Products Materials Included
 

Performance: Reproducibility

Highly Consistent ChIP of RNA Polymerase II by Multiple Researchers

The EpiTect Chip One-Day Kit Demonstrates a High Degree of Reproducibility. Four different researchers worked independently to collect duplicate samples of two million HCT116 cells. Each performed ChIP with the RNA Polymerase II antibody (RNA Pol II) or control IgG from a Epitect Chip Antibody Kit. The specific enrichment of ChIP DNA was analyzed by qPCR primers for the GAPDH proximal promoter. The results are expressed as individual percent of input results (insert table), their average (chart y-axis), and standard deviation (error bars). The figure shows that multiple researchers can achieve ChIP-qPCR of Pol II that agrees with only a 4.8 percent coefficient of variation.

Performance: Signal-to-Noise Ratios

Detecting RNA Polymerase II Enrichment on GAPD Promoter

The Epitect Chip System Detects RNA Polymerase II Enrichment at a Housekeeping Gene Promoter with Expected Signal-to-Noise Ratios. Triplicate wells of HCT-116 cells were fixed with formaldehyde. Chromatin was isolated and sonicated in preparation for immunoprecipitation using the Human RNA Polymerase II Antibody Kit and the One-Day Kit. The resulting enriched genomic DNA was then was purified and characterized by real-time PCR with primers for the proximal promoter region of the GAPD promoter and for an ORF-free region as a negative control. The results for each immunoprecipitation are expressed as percent of input, or the fraction of the total input DNA co-immunoprecipitating with RNA Polymerase II.

The immunoprecipitation (IP) with the RNA Polymerase II antibody (RNA Pol II) brings down much more GAPD proximal promoter than the IP with control IgG. Neither source of antibody precipitates a significant amount of a specific genomic DNA sequence within an ORF-free intergenic region or "promoter desert" lacking any known or predicted structural genes.

Performance: ChIP DNA Quality

Epitect Chip One-Day Kit Isolates DNA Free of PCR Inhibitors

ChIP One-Day Kit Isolates DNA Free of Inhibitors of Real-Time PCR. Chromatin was isolated from four million untreated HeLa cells and sheared by sonication. 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 ChIP One-Day Kit. Different volumes of each DNA preparation (1.0, 2.0, and 4.0 µL) were then used in appropriate ChIP-qPCR Primers to obtain threshold cycle values to generate a calibration curve.

The fit of the data to a straight line indicates 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 One-Day Kit, because the fold of enrichment is independent of the volumes of template from the immunopreciptated DNA.

 

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How It Works Performance (QC) Related Products Materials Included
 

Epitect Chip Antibody Kits
Validated high-quality ChIP-grade antibodies combined with positive and negative control PCR primer sets to insure ChIP performance.

EpiTect Chip qPCR Primers
Genome-wide qPCR primers specifically designed for analyzing genomic DNA enrichment from ChIP. The specificity and high amplification efficiencies are validated and guaranteed.

EpiTect Chip qPCR Array

Epitect Chip Customized PCR Array
Simultaneous analysis of multiple regulatory sites bound by transcription factors, co-regulators or modified histones. The two available array formats allow you to study:

  • Association of your transcription factors of interest with their binding sites on multiple genes of interest
  • Association of histone modification and chromatin remodelling with gene regulation across a 30-kb tiling region (-20 kb to +10 kb) around the TSS of any gene

RT² SYBR Green qPCR MasterMix
Optimized for the instrument in your lab by the experts in high-performance SYBR Green quantitative PCR

Epitect Chip qPCR Data Analysis Excel File
Excel-based template. Simply paste in your data, and results are automatically reported.

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How It Works Performance (QC) Related Products Materials Included
 

Please check the kit components immediately after you receive this package. We are not responsible for any missing items not reported within two (2) business days upon receipt.

Tube ID Contents Quantity
E01 10X PBS One 30-mL Bottle
PIC Protease Inhibitor Cocktail One 1.5-mL Clear-Capped Tube STORE AT -20 ºC!
E02 10X Stop Buffer One 15-mL Bottle
E03 IP Lysis Buffer One 15-mL Bottle
E04 IP Buffer A One 15-mL Bottle
E07 Protein A Beads One 2-mL Blue-Capped Tube CENTRIFUGE!
E21 IP Wash Buffer I One 15-mL Bottle
E22 IP Wash Buffer II One 15-mL Bottle
E23 IP Wash Buffer III One 15-mL Bottle
E24 IP Wash Buffer IV One 30-mL Bottle
E30 ChIP-Grade Proteinase K One 1.5-mL Red-Capped Tube STORE AT -20 ºC!
E31 DNA Extraction Beads One 2-mL Green-Capped Tube CENTRIFUGE!
E32 Elution Buffer Three 2-mL Yellow-Capped Tubes
E33 Column Binding Buffer One 15-mL Bottle
E34 Column Wash Buffer One 15-mL Bottle
DNA Spin Columns One Bag of 12 Columns
Non-Stick Elution Tubes (1.7-mL) Three Bags of 12 Tubes
2-mL Tubes (Sonication) One Bag of 6 Tubes
5-mL Tubes (Pre-Clear) One Bag of 6 Tubes

Storage Conditions

The Epitect Chip One-DAY Kit is shipped with ice packs.
The Protease Inhibitor Cocktail (PIC) and ChIP-Grade Proteinase K (E30) must be stored at -20 ºC. Centrifuge the Protein A Beads (E07) and DNA Extraction Beads (E31) at 4000 x g for 1 min at 4 ºC and return to its original position in the box. All other components must be stored at 4 ºC. We suggest that all other kit contents be kept in their original container to insure that no components are misplaced.

Shelf Life

All reagents are stable for 6 months after receipt of the kit if stored at the recommended temperature.

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