Genome-wide mapping of protein-DNA interactions is essential for a complete understanding of gene regulation. Mapping epigenetic marks and transcription factors is necessary to understand the regulatory networks controlling gene expression in a variety of biological systems.
The most widely used tool for examining these interactions is chromatin immunoprecipitation (ChIP) followed by massively parallel sequencing (ChIP-seq) (fig. 1). In a ChIP assay, DNA-protein complexes called chromatin are fixed to preserve the interactions. The chromatin is then extracted and sheared into small fragments.
The fragments are selectively immunoprecipitated using antibodies directed against the protein of interest and afterwards DNA and protein are separated. Via (rt)PCR, hybridization on microarrays, or direct sequencing the DNA fragments of defined sequence can be identified.
Recent advances have identified that that the interplay between chromatin and transcription is much more dynamic and complex than initially anticipated. ChIP-seq presents a needed enhancement of the ChIP-technology. Within one run up to hundreds of millions of DNA fragments can be sequenced producing fewer artefacts.
The short reads generated by next-generation sequencing (NGS) platforms are ideal for ChIP-seq and allow precise mapping of protein binding sites as well as improved identification of sequence motifs. With ChIP-seq you are able to investigate post translational modifications of chromatin and histone variants as well as nucleosome positioning. ChIP-seq is set to become the leading technology for genome-scale analysis of protein-DNA interactions benefiting from growing performance of sequencing platforms.
Chromatrap®: High-quality ChIP & ChIP-seq kits
Chromatrap® has established themselves as one of the leading ChIP and ChIP-seq kit distributors.
They were originally formed by Porvair Sciences in collaboration with Swansea University. In an academic/industrial partnership, the product range was increased inventing Chromatrap®, provides a revolutionary solid state platform for chromatin immunoprecipitation (ChIP) using spin columns or filter plates, amongst others.
Chromatrap® protocols offer a variety of advantages. It was developed to offer a faster, more sensitive method of chromatin immunoprecipitation. By avoiding the use of magnetic or agarose beads, non-specific background is eliminated together with reduced manual handling errors and subsequent sample loss.
Optimized elution buffer chemistry allows samples to be analyzed directly in qPCR without the need for DNA clean up, further reducing sample loss. Protein A or Protein G attached to inert, porous polymer increase the capture efficiency (fig. 2).
Chromatrap® utilizes the solid state technology in parallel with high throughput sequencing to deliver a precise ChIP-seq protocol from small cell numbers and chromatin concentrations (1-50 μg). Specifically adapted for greater chromatin concentrations, Chromatrap® ChIP-seq now combines the dynamic range of Chromatrap® with the downstream analysis power of deep sequencing, allowing the genome wide identification of transcription factor binding sites and specific DNA associated protein modifications with no limitation in scale and resolution.
Chromatrap® ChIP-seq products
Discover the wide range of Chromatrap® kitsSee all products
Explore now our stock of significant Chromatrap® products to get a complete set of tools to assist with a successful ChIP assay. The kits stand out with unique features such as enzymatic (ABIN3071686) or sonic (ABIN3071685) shearing or are designed for special template, e.g.:
Native ChIP-seq Protein A Kit (ABIN3071683)
The Chromatrap® Native ChIP kit which biologically shear the DNA of the chromatin by enzyme digestion. In native chromatin the proteins and histones wrapped around DNA are naturally linked and the traditional methods of shearing can easily disrupt the DNA- protein complexes. Native ChIP is mainly used for the study of histone modifiers and certain abundant transcription factors that are likely to be bound to DNA.Product Details
Premium Enzymatic ChIP qPCR Protein A (ABIN3071669)
Chromatrap® ChIP kits for qPCR rapidly improve ChIP using their unique patented technology streamlining the process, ensuring chromatin preparation to qPCR analysis can be performed in under 5 hours. The ChIP qPCR assay is a short convenient assay for low chromatin loading with very small slurry volumes (40 μLs) with no DNA clean up steps prior to qPCR.Product Details
FFPE ChIP-seq Protein A (ABIN3071673)
FFPE-ChIP utilizes chromatin extracted from Formalin Fixed Paraffin Embedded (FFPE) tissue or difficult to lyse samples. Tissues, such as biopsies, are routinely fixed in high formaldehyde concentration and embedded in paraffin in hospitals under non standardized conditions to allow long term storage without excessive sample degradation.Product Details
Chromatrap® ChIP qPCR Assay Kit - A Video Protocol
Chromatrap® Chromatin Immunoprecipitation qPCR Assay Kit - Video Protocol shows step by step how to perform a Chromatin IP from fresh human cells. Source: Chromotrap, Youtube