Characterization
Description(s):Transcriptional co-regulators interact specifically and non-covalently with one or multiple DNA-binding transcription factors (TFs) to either activate or repress the transcription of specific genes. Dysregulated expression of coregulator, such as SRC-1/NcoA-1, p300, CBP and HADC, has a significant effect on the regulation of a TF in the recruitment of other transcription factors and chromatin remodeling. For example, the upregulation of SRC-1 has known roles in mediating steroid receptor transcription in androgen receptor activity as well as prostate cancer progression.
Signosis has developed TF-Coregulator Interaction Plate Array Assay II allowing for high throughput studying of co-regulator interaction networks with 96 different TFs.
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-Coregulator Interaction Plate Array can simultaneously profile the transcriptional interaction of multiple TFs with a 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 to their corresponding TF. The 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 tube. 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 pre-coat plate and detected with streptavidin-HRP and substrate. The detected signals reflect the interacting TFs with the particular co-regulator of interest. Luminescence is reported as relative light units (RLUs) on a microplate luminometer.
