quantitative binding assays
Active Motif performs custom ChIP-qPCR assays to provide quantitative data on transcription factor binding and histone modification occupancy. Our Epigenetic Services team performs thousands of ChIP reactions every year. In order to make all of our experiments comparable and establish criteria for success, we have devised a unique normalization strategy. This normalization strategy, along with special attention dedicated to evaluating signal intensity and background levels, results in the highest quality ChIP data available. For an explanation of our normalization strategy, please download the ChIP-qPCR Data Normalization document.
The ChIP-qPCR Service includes:
Customers submit frozen tissues or cell pellets, then we:
- Prepare chromatin samples and sonicate.
- Perform ChIP with a ChIP-qualified antibody.
- Design and synthesize primers.
- Perform qPCR reactions in triplicate for the customer-selected binding sites and positive & negative control regions.
- Perform control qPCR reactions in triplicate using input DNA (unprecipitated genomic DNA).
- Analyze the data and deliver it to the customer.
The following papers cite the use of and/or provide additional information about ChIP-qPCR Services provided by Active Motif’s Epigenetic Services:
- “Targeting MYC dependence in cancer by inhibiting BET bromodomains” by Mertz et al (2011) Proc. Natl. Acad. Sci. USA 108(40):16669-16674.
- “The aryl hydrocarbon receptor interacts with c-Maf to promote the differentiation of type 1 regulatory T cells induced by IL-27” by Apetoh et al (2010) Nature Immunology 11(9):854-862.
- “Pet-1 is required across different stages of life to regulate serotonergic function” by Liu et al (2011) Nature Neuroscience 13(10):1190-1198.
- “Characterization of a Novel Small Molecule Subtype Specific Estrogen-Related Receptor α Antagonist in MCF-7 Breast Cancer Cells” by Chisamore et al (2009) PLoS 4(5):e5624.
- “Down-regulation of Death-associated Protein Kinase-2 Is Required for β-Catenin-induced Anoikis Resistance of Malignant Epithelial Cells” by Li et al (2009) JBC 284(4):2012-2022.