EpiTect ChIP qPCR ArraysFor application-focused quantitative detection of gene panels in chromatin immunoprecipitation experiments
EpiTect ChIP qPCR Arrays enable the simultaneous analysis of DNA–protein interactions using chromatin immunoprecipitation (ChIP)-enriched genomic DNA across a focused panel of genes. The carefully selected gene panels included in the EpiTect ChIP qPCR Arrays represent an epigenetically regulated biological pathway or disease state.
Performance
EpiTect ChIP qPCR Arrays provide high sensitivity, specificity, and reproducibility using SYBR® Green-based real-time PCR technology. ReproducibilityThe 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 verification of every EpiTect ChIP qPCR Assay, and high performance RT2 SYBR® Green Mastermix ensures the superior performance of EpiTect ChIP qPCR Arrays. See figure "Uniform amplification efficiency and specific PCR detection".
Principle
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. Available array formatsEpiTect ChIP qPCR Arrays are available in the following formats:
Procedure
Simply separately mix the input DNA fraction and specific antibody and control IgG ChIP DNA fractions with the appropriate ready-to-use RT2 SYBR Green qPCR Mastermix, aliquot equal volumes of each fraction into each well of its own plate, and then run the recommended real-time PCR cycling program.
Applications
EpiTect ChIP qPCR Arrays can be used for research into cancer, immunology, stem cells, toxicology, and biomarker discovery and verification, and are also powerful tools for studying regulatory mechanisms behind the gene expression changes observed with RT² Profiler PCR Arrays and Assays. EpiTect ChIP qPCR Arrays provide a reliable tool for the analysis of a panel of ChIP-enriched genomic sequences associated with transcription factors and co-regulators, modified and unmodified histones, and other DNA-binding proteins. EpiTect ChIP qPCR Arrays also streamline:
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