TF Reporter Plate Array I

Transactivation of a transcription factor (TF) is often measured by a reporter construct which contains a cis-element (DNA binding sequence) of the TF and a reporter gene such as a luciferase gene. When the reporter is delivered into cells, the activated TF binds to the cis-element and mediates the induction of the reporter gene. The induction can be measured via biochemical analysis of the reporter gene product. However, it is a single assay, one TF at a time. As any cellular change could result in the activation of multiple TFs, monitoring these TFs need multiple transfections and biochemical measurements. In addition, normalization is needed in order to normalize the transfection efficiencies. Signosis has developed a proprietary technology for monitoring the activation of multiple TFs simultaneously. In TF reporter plate array I, seven TF reporter vectors are mixed for monitoring the activation of seven TFs simultaneously, including NFkB, HIF-1, AP1, ISRE, Stat1, Stat3, and p53, along with a control TA vector without cis-element.

A. Benefits:

Based on its proprietary technology, Signosis developed TF reporter plate array, which offers following benefits:

  • Cell-based assay - It is a cell-based assay for monitoring the activities of TFs.

  • Multiplex - A single strip allows the measurement of the activities of seven TFs .

  • Quantitative comparison – The difference of two or more samples can be quantitatively analyzed.

B. Principle of the technology

Signosis developed a proprietary technology for monitoring the activation of multiple TFs simultaneously. In the technology, a series of reporter constructs are made, each of which contains a cis-element and a reporter tag sequence to a specific TF. When the constructs are delivered as a library into cells, the activation of TFs will bind to its corresponding constructs and mediate the expression of the tag sequences. The expressed tag sequences are analyzed via a cDNA plate array, with which each specific tag sequence is analyzed by a corresponding complementary tag capture oligo immobilized onto an assigned well. In TF reporter plate array I, seven TF reporter vectors are mixed for monitoring the activation of seven TFs simultaneously, including NFkB, HIF-1, AP1, ISRE, Stat1, Stat3, and p53, along with a control TA vector without cis-element.

Figure 1
Diagram

Figure 2. TF reporter array analysis of the activities of seven transcription factors. HeLa cells were transfected with TF reporter plasmids without (purple) and with 20ng/ul TNF (red) or 15% serum treatment (yellow) for 4 hours. Total RNAs were prepared. RNAs were subjected to TF reporter plate array.

 

 Product Size Cat. Price
TF Reporter Plate Array I 6 AP-2101 $489
  • TF Reporter Plate Array (PDF)

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