EpiTect ChIP Custom qPCR Arrays
For quantitative detection of custom-defined gene panels in chromatin immunoprecipitation experiments
EpiTect ChIP Custom qPCR Arrays enable the simultaneous analysis of DNA–protein interactions using chromatin immunoprecipitation (ChIP)-enriched genomic DNA across a focused panel of genomic loci tailored to specific research interests. Analyze multiple gene-specific binding sites for the same transcription factor, the histone code at the proximal promoter of multiple genes, or the histone code or binding sites for multiple transcription factors across one entire gene promoter.
EpiTect ChIP qPCR Arrays provide high sensitivity, specificity, and reproducibility using SYBR® Green-based real-time PCR technology.
The complete EpiTect ChIP qPCR Array system demonstrates a high degree of reproducibility across technical replicates, lots, and instruments. This consistency ensures reliable detection of differences in genomic DNA enrichment among biological samples. See figures "Consistent intra- and inter-plate performance" and "Consistent performance with different amounts of DNA, instruments, or handling conditions".
Specific and accurate ChIP-qPCR detection
One prerequisite for ChIP qPCR array technology is uniform and high PCR amplification efficiency, which allows a reciprocal comparison of ChIP enrichment among all genomic loci analyzed. The unique combination of a proprietary ChIP-PCR primer design algorithm, rigorous experimental validation of every EpiTect ChIP qPCR Assay, and high performance SYBR® Green Mastermix ensures the superior performance of EpiTect ChIP qPCR Arrays. See figure "Uniform amplification efficiency and specific PCR detection".
Analyzing the histone code across an entire gene promoter
One application of EpiTect ChIP Custom qPCR Arrays is the monitoring of differential histone modifications across a gene. The EpiTect ChIP Custom qPCR Array quickly maps histone modifications surrounding the transcription start site (TSS) of the CDKN1A gene (see figure "Monitoring of differential histone modifications").
Regulating gene expression requires dynamic but regulated interactions between genomic DNA and proteins, such as transcription factors, coactivators, corepressors and modified histones. One important technique for studying the discovery of new and the timing and extent of known in vivo protein-DNA binding is chromatin immunoprecipitation (ChIP). Chemical crosslinking at the end of a biological experiment covalently captures and freezes the protein–DNA interactions. Sonication shears the chromatin into manageable sizes for the sensitive detection of specific genomic DNA sequences. Standard immunoprecipitation pulls down a target protein of interest and its bound DNA. Crosslink reversal and DNA purification, followed by real-time PCR detection of specific sequences then quantifies the amount of protein-bound DNA.
The EpiTect ChIP Custom qPCR Array contains a set of optimized real-time PCR primer assays on 96-well or 384-well plates for analysis of in vivo protein–DNA interactions on custom-defined gene content tailored to a specific research interest. The EpiTect ChIP qPCR Assays used in the EpiTect ChIP Custom qPCR Arrays are genomewide, laboratory-verified, real-time PCR tools to analyze any promoter region in human, mouse, and rat samples. The EpiTect ChIP Custom qPCR Array performs ChIP DNA analysis with real-time PCR sensitivity and the multi-genomic loci profiling capability of a ChIP-on-chip. It allows the simultaneous analysis of multiple genomic loci and samples in 96- or 384- well format on any real-time PCR instrument. The performance of the EpiTect ChIP Custom qPCR Arrays is guaranteed when used with the appropriate RT2 SYBR Green qPCR Mastermix.
Fully customizable arrays
EpiTect ChIP Custom qPCR Arrays are available in a series of 96- and 384-well formats (see figure "EpiTect ChIP Custom qPCR Array formats"). Different formats are also possible. Please inquire for more information and to place an order. EpiTect ChIP Custom qPCR Arrays are manufactured, laboratory-verified, and shipped in 2–3 weeks from order verification.
Separately mix input DNA fractions and specific antibody and control IgG ChIP DNA samples with the appropriate ready-to-use PCR Mastermix, aliquot equal volumes to the correct and designated wells of the same plate or into each well of a fraction’s own plate, and then run the recommended real-time PCR cycling program.
Data analysis is based on the ∆CT method to calculate percent enrichment with normalization of the specific antibody and control IgG ChIP DNA fraction to the input DNA fraction. An easy-to-use Excel-based data analysis template for these calculations can be tailor-made.
EpiTect ChIP qPCR Arrays can be used for research into cancer, immunology, stem cells, toxicology, and biomarker discovery and validation, and are also powerful tools for studying regulatory mechanisms behind the gene expression changes observed with RT² Profiler PCR Arrays and Assays. EpiTect ChIP Custom qPCR Arrays provide a reliable tool for the analysis of a panel of ChIP-enriched genomic sequences associated with transcription factors and coregulators, modified an unmodified histones, and other DNA-binding proteins.
EpiTect ChIP Custom qPCR Arrays also streamline:
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