Cell Signaling HaloCHIP™ System Product HaloCHIP™ System For Research Use Only. Not for Use in Diagnostic Procedures. Description: The HaloCHIP™ System is a novel method designed for the covalent capture of intracellular protein:DNA complexes without the use of antibodies and offers an efficient and robust alternative to the standard chromatin immunoprecipitation (ChIP) method. Proteins of interest are expressed in cells as HaloTag® fusion proteins, crosslinked to DNA with formaldehyde and then captured on HaloLink™ Resin, which forms a highly specific, covalent interaction with the HaloTag® portion of the fusion protein. Stringent washing removes nonspecific proteins and DNA, and heating reverses the crosslinks between the DNA and the fusion protein and releases the captured DNA fragment, which subsequently can be purified. Technology Size Cat.# Price (Fr) 20 reactions G9410 783.00 Features: • No Requirement for Antibody: No need to make your own or purchase expensive, qualified antibodies. • Obtain Results Faster: Obtain data in 24–48 hours with fewer steps to minimize potential experimental errors. • Improved Signal-to-Noise Ratios: Enables detection of small changes in protein binding patterns using a minimal number of cells. Storage Conditions: The TE Buffer (pH 8.0), Reversal Buffer and NucleaseFree Water may be stored at room temperature. Store the HaloLink™ Resin, Mammalian Lysis Buffer and High Salt Wash Buffer at 4°C. Store the HaloCHIP™ Blocking Ligand at –20°C. Protocol HaloCHIP™ System Technical Manual Part# TM075 14 Covalent Capture of Chromatin Complexes Using HaloTag® Transfection fusion construct HaloTag® TF HT TF HaloCHIP™ Blocking Ligand Experimental Sample HaloLink™ HT TF HaloLink™ Block HaloTag binding HT Untransfected Cells OR Controls Split Sample TF HT TF HaloLink™ Wash HaloLink™ Resin Covalent capture allows highly stringent washes to remove nonspecific proteins and DNA. Sample DNA Background DNA Release of DNA by reversal of crosslinks Protein remains bound to the HaloLink™ Resin. Analyze Analyze TF HT Schematic diagram of the HaloCHIP™ System. = HaloTag® protein = Transcription factor Add HaloCHIP™ Blocking Ligand to the control sample to prevent binding to HaloLink™ Resin. Capture using the HaloLink™ Resin HT Formaldehyde Crosslinking Lysis, Sonication TF HT Expression of HaloTag® Fusion Protein For complete and up-to-date product information visit: www.promega.com/catalog 251 Protein Purification and Interactions 1–1.5 days 6633MB Pagina 254
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