AP1 Filter Plate assay

AP-1, a dimeric protein complex composed of Jun and Fos proteins, plays a key role in the control of cell death, proliferation and transformation. Ap1 protein can be activated by many stimuli, including TPA, serum, growth factors and oncoproteins, TNF and IL-1, as well as by a variety of environmental stresses, such as UV radiation. When it is activated, AP1 protein binds to AP- DNA recognition element within the promoter regions of target genes and regulates the gene expression. Signosis has developed AP1 filter assay for monitoring DNA binding activity of AP1, which can be used for analyzing multiple samples simultaneously.

A. Benefits:

Signosis proprietary AP1 filter plate assay offers following benefits:

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

• High throughput screen- 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

Signosis’ AP1 filter assay is a plate-based analysis for monitoring the activity of AP1. In the assay, biotin labeled AP1 DNA binding sequence is mixed with nuclear extract to allow formation of AP1-DNA complex. A filter plate is used to retain bound AP1 probe and remove free DNA probe. The bound pre-labeled AP1 probe is then eluted from the filter and hybridize to the corresponding well of 96 well of Hybridization Plate for quantitative analysis. The captured AP1 probe is further detected with streptavidin-HRP. Luminescence is reported as relative light units (RLUs) on a microplate luminometer.