Characterization
Description(s):Transcription factors (TFs) are a group of cellular proteins that play essential roles in regulating gene expression. They act as sensors to monitor cellular change and convert the signals into gene expression. Often, TFs regulate gene expression not only through directly binding to their DNA binding site on the target promoter region, but also by interacting with other transcription factors or regulators. Since changes in transcriptional interaction patterns can have dramatic effects on gene expression and cellular responses to stimuli, monitoring multiple TF/TF and TF/co-regulator interactions simultaneously could yield a greater understanding of regulatory mechanisms. Signosis’ TF Protein Interaction Plate Array II can monitor TF/TF or TF/co-regulator interactions among 96 different TFs simultaneously in relation to one TF or co-regulator of interest.
Applicable Grid:
| 1 | 2 | 3 |
4 |
5 |
6 |
7 | 8 | 9 | 10 |
11 |
12 | |
| A | AP1 | CDP | GATA |
NF-1 |
Pit |
Stat3 |
XBP | FOXG1 | HoxA-5 | NRF2(A) |
ProX1 |
SOX2 |
| B | AP2 | CREB | GR/PR | NFAT | PPAR | Stat4 | AP3 | FOXO1 | HSF | Oct-1 | RB | SOX9 |
| C | AR | E2F-1 |
HIF |
NF-E2 | PXR | Stat5 | AP4 | FREAC-2 | KLF4 | Pax2 | RUNX | SOX18 |
| D | ATF2 | EGR | HNF4 | NFkB | SMAD | Stat6 | COUP-TF | Gli-1 | MyoD | Pax3 | ROR(RZR) | SRY |
| E | Brn-3 | ER | IRF | OCT4 | Sp1 | TCF/LEF | ELK | Gfi-1 | MZF | Pax8 | RXR | TFE3 |
| F | C\EBP | Ets | MEF2 | p53 | SRF | YY1 | FOXA1 | HEN (NSCL-1) | Nkx2-5 | PIT1 | SF-1 | USF-1 |
| G | CAR | FAST-1 | Myb | Pax-5 | SATB1 | TR | FOXC1 | HNF-1 | Nkx3-2 | PLAG1 |
SMUC |
VDR |
| H | CBF | GAS/ISRE | Myc-Max | Pbx1 | Stat1 | TFIID | FOXD3 | HOX4C | NRF1 | MEF1 | Snail | WT1 |
Principle
Signosis’ TF-TF Interaction Plate Array can simultaneously profile the transcriptional interaction of multiple TFs with a TF or co-regulator of interest. In this assay, a series of unique biotin-labeled probes are provided that correspond with the consensus sequences of individual TF DNA-binding sites. Therefore, each probe represents an individual TF. When the probe mix is incubated with nuclear extract, individual probes bind their corresponding TF. The TF or co-regulator of interest is then immunoprecipitated, along with transcriptionally interacting TFs, using a corresponding antibody and protein G or A agarose beads in a column. Unbound probes and proteins are washed away. The bound probes are then detached from the complex and are subsequently denatured. The biotin-labeled DNA strands are hybridized on a white plate and detected with streptavidin-HRP and substrate. The detected signals reflect the interacting TFs with the particular TF or co-regulator. Luminescence is reported as relative light units (RLUs) on a microplate luminometer.
