Enrichment of Highly Abundant Epigenetic Marks from Endometrial Stromal Cells with Porvair’s Chromatrap®

30 Apr 2014
Sarah Thomas
Associate Editor

Product news

Chromatrap®, the novel solid-based matrix for chromatin immunoprecipitation (ChIP) assays, available from Porvair Sciences, has been successfully used to isolate high quality chromatin from difficult biopsy material.

Focusing on the precipitation of epigenetic marks from primary endometrial stromal cells, researchers have produced excellent enrichment of high abundant epigenetic marks from 1 million cells isolated from a tissue biopsy. The selective enrichment using Chromatrap was highlighted by the selective amplification of both positive and negative gene targets relative to negative IgG. Further the work described demonstrates how the Chromatrap ChIP Assay Kit is able to delineate the balance between active and repressive associated marks and also is compatible with human tissue.

The protocol to enrich epigenetic histone marks from primary human tissue cells is detailed in an application note. Click on the company website link to download the application note.

Chromatrap® uses a solid phase porous polymer functionalised with protein A or G, allowing much greater chromatin and antibody capturet than bead based methods. The Chromatrap spin column approach offers significant advantages compared to traditional methods based on sepharose or magnetic beads, which involve many steps of separation, pipetting and re-suspension. Independent tests with Chromatrap demonstrate the reproducibility of the assay and the strong fold amplification above negative IgG background, enables the targeting of both abundant and low abundant marks at multiple gene targets. Levels of DNA pull down are up to 25 times higher than with traditional bead based procedures and top performance even with samples as small as 50ng-3000ng chromatin per immunoprecipitation enables Chromatrap to be used on small scale biopsies .

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Genome AnalysisGenomics, the study of genomes, includes functional genomics, evolutionary genomics and comparative genomics. There are many genomic technologies such as DNA sequencing of whole genomes, computational biology and bioinformatics. DNA and nucleic acids must be isolated and concentrated from cells for analysis with kits, automated analyzers and software. Other useful technologies for studying genomics include PCR, microarrays and electrophoresis.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.Epigenetic AnalysisEpigenetic analysis products are used to study heritable phenotype changes that do not alter the DNA sequence, such as DNA modifications affecting gene activity. Determine whether genes have been methylated with DNA methylation quantification kits or methylation microarrays. Assess epigenetic regulation with histone modification kits or ChIP-on-chip microarrays. Kits are also available for bisulfite conversion and amplification of your samples, as well as epigenomic library preparation kits for next-generation sequencing (NGS). Find the best epigenetic products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ChIP-Sequencing