EpiTect ChIP qPCR Assays
For quantitative detection of interactions between DNA and nuclear proteins in chromatin immunoprecipitation experiments
EpiTect ChIP qPCR Assays provide experimentally verified genomewide real-time PCR primers specifically designed for the analysis of ChIP-enriched genomic DNA. They focus on specific 1 kb genomic regions in gene promoters, allowing detailed analysis of the dynamic interaction between DNA and nuclear proteins. Assays can be selected by gene, transcription factor, or chromosomal position and are available for human, mouse, and rat.
Regulating gene expression requires dynamic, but regulated, interactions between genomic DNA and proteins, such as transcription factors, co-activators, co-repressors, 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 cross-linking at the end of a biological experiment covalently captures and freezes the protein-DNA interactions. Sonication shears the chromatin into manageable sizes for sensitive detection of specific genomic DNA sequences. Standard immunoprecipitation pulls down a target protein of interest and its bound DNA. Cross-link reversal and DNA purification followed by real-time PCR detection of specific sequences then quantifies the amount of protein-bound DNA.
The target-sequence–specific EpiTect ChIP qPCR Assays are designed with a proprietary algorithm developed specifically to address the challenges presented by genomic DNA sequences. Each assay is also experimentally verified for the high amplification efficiency and specificity expected of optimal SYBR® Green-based real-time PCR assays. The performance of the EpiTect ChIP qPCR Assays is guaranteed when used with the appropriate RT2 SYBR® Green qPCR Mastermix.
The EpiTect ChIP qPCR system provides a complete platform for the in vivo analysis of the dynamic interaction between DNA and nuclear proteins that play a central role in gene regulation. This technology integrates chromatin immunoprecipitation DNA preparation and real-time PCR analysis.
Simply set up separate PCR tubes with input DNA fraction, control ChIP DNA fraction, and all specific antibody ChIP DNA fractions with the appropriate RT2 SYBR Green qPCR Mastermix. Then, run the recommended 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 raw data. An easy-to-use Excel-based data analysis template for these calculations enables simple analysis.
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 qPCR Assays can be used for quantifying immunoprecipitated DNA following ChIP, and provide a reliable tool for the analysis of a panel of ChIP-enriched genomic sequences associated with transcription factors and coregulators, modified and unmodified histones, and other DNA-binding proteins.
EpiTect ChIP qPCR Assays also streamline:
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