In the HaloCHIP™ method, a HaloTag® fusion protein is expressed either transiently or stably in mammalian cells and crosslinked complexes are directly captured from the cellular lysate via covalent binding to a HaloTag®-specific resin. The covalent linkage allows for extensive washing to remove non-specific proteins and DNA, before reversal of the crosslinks to release the bound DNA fragments.
- Simple, 1-day ChIP protocol, without antibodies.
- Low Background. HaloTag® covalent capture allows stringent washing compared to antibody-dependent ChIP.
- Savings compared to conventional ChIP kits, which require the purchase of an untested antibody.
Similar fold enrichment pull-down results were obtained using HaloCHIP™ to pull-down CREB:DNA partners as compared to conventional CREB ChIP.
qPCR Data. Conventional (antibody-dependent) ChIP was compared to HaloCHIP for CREB:DNA binding sequences. No significant differences in enrichment patterns were found when comparing known CREB-binding sites: Fos, Jun, and p27.
There is excellent reproducibility in binding between HaloCHIP™ and conventional antibody-dependent ChIP assays.
ChIP-chip data. High-resolution analysis demonstrates nearly identical promoter enrichment profiles for CREB binding when comparing CREB-HaloCHIP™ to CREB-ChIP.
Hartzell, D.D. et al. (2009) functional analysis of the CREB signaling pathway using HaloCHIP-chip and high throughput reporter assays.
BMC Genomics., 2009; 10: 497.
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