Combine with TF-TF Interaction Array II and TF-TF Interaction Array III for a total of 244 unique TFs.- Novel—DNA array-based system for mapping the network of TF-TF interactions
- Quick—screen for 54 TF-TF interactions in one experiment
- Highly sensitive—HRP-based detection system
- Safe—no radioactivity required
TF-TF Interaction ArraysTF-TF Interaction Arrays enable you to determine how a particular TF interacts with multiple other TFs—all in one experiment. Unlike traditional methods, such as co-immunoprecipitation and super-gel shift, this array-based method generates a global view of TF-TF interactions. Three versions are currently available. TF-TF Interaction Array I detects the activity of 54 TFs while TF-TF Interaction Array II and TF-TF Interaction Array III detect the activity of 96 and 94 TFs, respectively. Together, these arrays can profile the activity of 244 unique TFs. TF-Specific TF Interaction Arrays for NFκB, PPAR, GR, p300, and SRCArrays for analysis of specific transcription factor complexes, focusing on interactions between specific TFs and other TFs. - Novel—DNA array-based system for mapping the network of NFκB, PPAR, GR, p300, and SRC-1 interactions
- Simple—for identification of NFκB, PPAR, GR, p300, and SRC-1 interaction changes
- Quick—screen for 54 or 96 TF interactions in one assay
- Safe and Sensitive—HRP-based chemiluminescent detection system
TF-Specific-TF Interaction Arrays enable you to determine how NFκB, PPAR, GR, p300, and SRC-1 interact with multiple TFs—all in one experiment. Unlike traditional methods, this array-based technique generates a global view of NFκB, PPAR, GR, p300, and SRC-1 protein interactions. And by profiling NFκB, PPAR, GR, p300, and SRC-1 and TF activity simultaneously, you can gain valuable insight into the signaling pathways through determining signal-associated alterations in TF-binding. The TF-TF Interaction Array uses a proprietary, patent-pending technology for detecting interactions between TFs. Instead of directly measuring interactions between proteins, this Array employs an exquisitely sensitive indirect approach. This technology is based on the fact that every TF has its own responsive DNA binding element, or cis-element. TF-TF interactions can be assessed by looking at the bound cis-elements—i.e., the interactions can be indirectly visualized when corresponding cis-elements are hybridized to the membrane. In this way, you can quickly see how one TF interacts with multiple other TFs. Individual results can then be confirmed by co-immunoprecipitation or super-gel shift. The TF-TF Interaction procedure is simple and straightforward: - First, incubate your nuclear extract with the provided set of biotin-labeled, double-stranded oligonucleotide probes, which represent a known library of cis-elements. During the incubation step, these TF probes bind to their specific TFs in your nuclear extract.
- The next step is immunoprecipitation: using a specific antibody against your TF of interest, pull out that TF and associated TFs, along with corresponding cis-elements.
- Next, wash away free cis-elements and nonspecific binding proteins.
- Finally, elute the cis-elements and hybridize them to the Array membrane (provided), which is spotted with different TF consensus sequences.
The Array detects TF interactionsThe figures below illustrate how the TF-TF Interaction Array can yield informative results. In Figures A and B, the Array was used to determine which TFs interact with p53. Protein pull-down assays (Figures C & D) then confirmed that CREB and PAX-5 interact with p53.  | TF-TF Interaction Array identifies p53 and associated transcription factors as described in literature. |  | Figure A: A431-PMA-treated nuclear extract was mixed with the Probe Mix. Immunoprecipitation was performed using a polyclonal p53 antibody, followed by "pull down" using Dynal Protein G beads. The beads and associated TF-probe complexes were washed four times and the probes were eluted. The eluted probes were then hybridized to the Protein/DNA Array membrane. The cis-elements that hybridized corresponded to TFs that interact with p53, as reported in the literature (1–8).
Figure B: A431-PMA-treated nuclear extract immunoprecipitated with normal IgG instead of the p53 antibody. |  | Co-immunoprecipitation of A431-PMA-treated nuclear extracts confirms that p53 interacts with CREB, as seen in Figure A. Nuclear extract alone (Lane 1) and two immunoprecipitates (Lane 2 immunoprecipitated with normal IgG; Lane 3 immunoprecipitated with p53 antibody) were run on SDS-PAGE. A Western blot was hybridized with monoclonal anti-CREB antibody (Santa Cruz Biotech) to confirm that p53 interacts with CREB. | TF-Specific Interaction Array for NFkBNuclear extracts from untreated or TNFα-treated HeLa cells were incubated with the NFκB Probe mix. Immunoprecipitation with a NFkB-specific antibody was performed, and the protein-bound probes were then isolated and hybridized to the NFκB-TF Interaction Array membrane. Chemiluminescent images were collected using a FluorChem imager (Alpha Innotech), revealing changes in TF binding to NFkB upon TNFα stimulation. User Manuals and Product Inserts:- TF Interaction Array User Manual PDF
- TF-TF Specific PD Array User Manual PDF
- Stripping Procedure PDF
- TF/TF Interaction Array I Product Insert PDF
- TF/TF Interaction I Refill Kit Product Insert PDF
- TF/TF Interaction Array II Product Insert PDF
- TF/TF Interaction II Refill Kit Product Insert PDF
- TF/TF Interaction Array III Product Insert PDF
- TF/TF Interaction III Refill Kit Product Insert PDF
Schematic Diagrams:- TF-TF Interaction Array Schematics PDF
Related Products:References:- Hiratochi, M. et al. The Delta intracellular domain mediates TGF-beta/Activin signaling through binding to Smads and has an important bi-directional function in the Notch-Delta signaling pathway. Nucleic Acids Res 35(3):912-22 (2007). PDF
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RNA in situ Analysis
