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. The transcriptional co-activator steroid receptor coactivator 1 (SRC-1) is a highly flexible nuclear receptor coregulator that regulates gene expression and can interact with many families of transcription factors, and is often found to be overexpressed in many cancers.
Signosis has developed TF-Coregulator SRC-1 Interaction Plate Array Assay I, allowing for high throughput studying of co-regulator interaction networks with 48 different TFs.
Applicable Grid:
|
|
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
|
A |
AP1 |
CDP |
GATA |
NF-1 |
Pit |
Stat3 |
AP1 |
CDP |
GATA |
NF-1 |
Pit |
Stat3 |
|
B |
AP2 |
CREB |
GR/PR |
NFAT |
PPAR |
Stat4 |
AP2 |
CREB |
GR/PR |
NFAT |
PPAR |
Stat4 |
|
C |
AR |
E2F-1 |
HIF |
NF-E2 |
PXR |
Stat5 |
AR |
E2F-1 |
HIF |
NF-E2 |
PXR |
Stat5 |
|
D |
ATF2 |
EGR |
HNF4 |
NFkB |
SMAD |
Stat6 |
ATF2 |
EGR |
HNF4 |
NFkB |
SMAD |
Stat6 |
|
E |
Brn-3 |
ER |
IRF |
4-Oct |
Sp1 |
TCF/LEF |
Brn-3 |
ER |
IRF |
4-Oct |
Sp1 |
TCF/LEF |
|
F |
C\EBP |
Ets |
MEF2 |
p53 |
SRF |
YY1 |
C\EBP |
Ets |
MEF2 |
p53 |
SRF |
YY1 |
|
G |
CAR |
FAST-1 |
Myb |
Pax-5 |
SATB1 |
TR |
CAR |
FAST-1 |
Myb |
Pax-5 |
SATB1 |
TR |
|
H |
CBF |
GAS/ISRE |
Myc-Max |
Pbx1 |
Stat1 |
TFIID |
CBF |
GAS/ISRE |
Myc-Max |
Pbx1 |
Stat1 |
TFIID
|
Principle
Signosis’ TF-Coregulator SRC-1 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.
