Instructions for Use of Product(s)
Literature # TM075
The HaloCHIP™ System is a novel system that covalently captures crosslinked protein:DNA complexes from cells without the need for antibodies, providing a robust and efficient alternative to the standard chromatin immunoprecipitation (ChIP) method. The HaloCHIP™ System takes advantage of the HaloTag® protein, which can be fused to any protein and mediates a covalent interaction with various ligands, including a resin-based ligand, HaloLink™ Resin. For this application, a DNA-binding protein of interest is fused to the HaloTag® protein via cloning in a HaloTag® Vector. The recombinant vector construct is transfected into cells for either stable or transient expression. Then the cells are treated with formaldehyde to induce covalent protein:DNA crosslinks, lysed and sonicated to shear the chromatin into smaller fragments for purification by the HaloCHIP™ System. Using the HaloCHIP™ System, these crosslinked complexes can be directly captured from the lysate through covalent binding between the HaloTag® moiety of the fusion protein and the HaloLink™ Resin. Stringent washing removes nonspecific proteins and DNA. Subsequent heating of the HaloTag® fusion protein:DNA complexes on the resin reverses the crosslinks between the DNA and the fusion protein and releases the captured DNA fragments.
Summary of Changes, 12/15:
The following change was made to the 12/15 version of this technical manual: Patent statements were updated.
Printed in USA. Revised 12/15.