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The Active Motif Epigenetic Services team performs thousands of ChIP reactions every year. Each experiment is unique, using different amounts of chromatin, different antibodies, different primers with varying efficiencies and different final volumes (depending on the downstream application). In order to make all of our experiments comparable and establish criteria for what is “normal”, we have devised a normalization strategy that takes all of these variables into account and presents the data as Binding Events Detected per 1000 cells. This unique normalization strategy enables us to directly compare every experiment we perform. By having acceptable ranges for signal intensity and background levels, we can easily determine if ChIP reactions are successful and identify outlying reactions that may need to be repeated. For an explanation of our normalization strategy, please download the ChIP-qPCR Data Normalization document.
Our sensitive ChIP-qPCR assays are quantitative and can reproducibly detect changes in transcription factor binding as low as 1.25 fold.
The ChIP-qPCR Service includes
The customer submits 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 of the customer-selected binding sites as well as 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.
To learn more, please give us a call or send us an Epigenetic Services Information Request. You can also download Active Motif’s Epigenetic Services Profile.
ChIP-qPCR normalization.
Active Motif has developed a unique ChIP-qPCR normalization strategy that takes into account the starting amount of chromatin, final ChIP volume, and primer efficiency. This approach allows all ChIP reactions to be presented on the same scale; negative control primers typically give a signal below 1; enrichment of specific sites can be between 10 and 500.
| Name | Cat No. | Price | |
|---|---|---|---|
| FactorPath™ ChIP-qPCR | 25004 | Request Quote | |
| HistonePath™ ChIP-qPCR | 25014 | Request Quote | |
| Active Motif Epigenetic Services Profile |
| Epigenetic Services Cell Fixation Protocol |
| Epigenetic Services Sample Submission Form |
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.
