Product Name Catalog # Price   Qty
TF-TF Interaction Plate Array I FA-3001 $529
- +

Characterization

Description:

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 I can monitor TF/TF or TF/co-regulator interactions among 48 different TFs simultaneously in relation to one TF or co-regulator of interest.


Applicable Grid:


List of Applicable TFs:

  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 OCT4 Sp1 TCF/LEF Brn-3 ER IRF OCT4 Sp1 TCF/LEF
F C\EBP Ets MEF2 p53 SRF TR C\EBP Ets MEF2 p53 SRF TR
G CAR FAST-1 Myb Pax-5 SATB1 YY1 CAR FAST-1 Myb Pax-5

SATB1

YY1
H CBF GAS/ISRE Myc-Max Pbx1 Stat1 TFIID CBF GAS/ISRE Myc-Max Pbx1 Stat1 TEIID

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.

Data

 

tf tf

Analysis of NFkB interaction patterns in HeLa and TNFalpha-treated HeLa. The cells were treated with and without TNFalpha for 1 hour. The nuclear extracts were prepared and incubated with cis-element probe mix and antibody against NFkBp65. The NFkB interacting TFs were pulled down and the bound probes were eluted and hybridized to the plate.

Literature

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