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 p300 is a histone acetyltransferase that is typically recruited to transcriptional enhancer and interacts with transcription factors and regulates gene expression.
Signosis has developed TF-Coregulator p300 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 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 p300. Luminescence is reported as relative light units (RLUs) on a microplate luminometer.
