2012 Promega catalogue

Cell Signaling
HaloCHIP System
Product Size Cat.# Price ($)
HaloCHIP System 20 reactions G9410 829.00
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.
HaloTag ®
fusion construct
Transfection
HT
TF
HaloLink
HaloLink
Covalent Capture of Chromatin Complexes
Using HaloTag ® Technology
Experimental
Sample
HT
HT
TF
TF
Schematic diagram of the HaloCHIP System.
For complete and up-to-date product information visit: www.promega.com/catalog
TF
HT
TF
HT
HaloCHIP
Blocking Ligand
Controls
Untransfected Cells
OR
Block HaloTag binding
HaloLink
HT
Sample DNA Background DNA
Analyze Analyze
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 Nuclease-
Free 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 Part#
HaloCHIP System Technical Manual TM075
TF
Expression of HaloTag ®
Fusion Protein
Formaldehyde Crosslinking
Lysis, Sonication
Split Sample
Add HaloCHIP Blocking
Ligand to the control sample
to prevent binding to
HaloLink Resin.
Capture using the
HaloLink Resin
Wash HaloLink Resin
Covalent capture allows
highly stringent washes
to remove nonspecific
proteins and DNA.
Release of DNA by reversal
of crosslinks
Protein remains bound
to the HaloLink Resin.
HT
TF
= HaloTag ® protein
= Transcription factor
1–1.5 days
6633MB
249
14
Protein Purification and Interactions
Section
Contents
Table of
Contents