The chromatin immunoprecipitation (ChIP) assay is a major tool in the study of genomic processes and gene regulation in vivo. It is widely known control gene expression involves multiple proteins and great number of their modifications. Chip assay is conventionally performed in test tubes one at a time, which limits the ability to gain comprehensive picture of the complex genomic events. To simplify the assay in high throughput way, Signosis has developed a 96 plate based ChIP assay, where all of steps from IP to DNA elution are handled in microplate wells without sample transfers in a high throughput way. The assay is an ideal tool for studying up to 16 protein-DNA bindings in one sample or one protein-DNA binding in up to 16 samples, or multiple protein-DNA bindings in multiple samples/treatments.
This assay utilizes the surface-immobilized antibodies two 8-well strips. The entire procedure from chromatin precipitation to PCR-ready DNA is done on the same wells without sample transfers. The microplate is first blocked and coated with protein A&G, The plate is then coated with one or more antibodies of interest. The cells in 96-well culture plate or any size of dishes are cross-linked. After chromatin fragmentation, the cell lysates are transferred and incubated on the antibody-coated plate to form antibody-protein-DNA complex. After several wash steps, the bound DNAs are subsequently eluted. The eluted DNAs can be used for PCR analysis or next generation sequencing. The microplate-based ChIP assay has several important advantages over the tube-based assay: simple sample handling, high throughput, improved sensitivity and reproducibility.