TF Filter Plate Assays

Transcription factors (TFs) are a group of cellular proteins that play essential roles in regulating gene expression. They act as a sensor to monitor cellular change and convert the signals into gene expression. A specific TF is often regulated by multiple upstream proteins. Monitoring the activation of a TF is critical to uncovering the mechanism of signal pathways and cellular regulations. Signosis developed 48 different filter assays for monitoring the activities of TFs. Unlike gel shift assays, 96 samples can be analyzed in one experiment.

A. Benefits:

Signosis' proprietary TF filter plate assays offer following benefits:

• Simple and quick -It is simpler and quicker than gel shift assay.

• High throughput screen- A specific TF in multiple samples can be analyzed in one single assay.

• Cost-effective detection- The cost is lower than ELISA-based TF assays.

• Quantitative comparison The difference of two or more samples in TF activity can be quantitatively analyzed.

B. Principle of the technology

In Signosis' TF filter plate assays, a specific biotin-labeled TF DNA binding sequence is mixed with nuclear extracts to allow formation of TF-DNA complexes. A filter plate is used to retain bound DNA probe and remove free probe. The bound pre-labeled DNA probe is then eluted from the filter and collected for quantitative analysis through DNA plate hybridization. The captured DNA probe is further detected with streptavidin-HRP. Luminescence is reported as relative light units (RLUs) on a microplate luminometer.