Principle 

The NFκB cDNA Plate Array Assay is designed to analyze the expression of over 20 NFκB-regulated genes directly from cell lysates, without the need for RNA extraction. This is made possible by Signosis’ proprietary CL™ cDNA synthesis technology, which enables reverse transcription to be performed directly in crude cell lysates.

1. Cell Lysis & cDNA Synthesis
   Cells are lysed, and reverse transcription is performed directly in the lysate using the CL™ cDNA Synthesis Kit, eliminating the need for RNA isolation.

2. cDNA Hybridization
   The resulting biotin-labeled cDNA probes are added to a 96-well plate, where each well is pre-coated with gene-specific oligonucleotides targeting NFκB-regulated genes.

3. Detection
   Captured cDNA is detected using streptavidin-HRP and a chemiluminescent substrate. The signal is read on a microplate luminometer and reported in relative light units (RLUs).

4. Data Interpretation
   The luminescent signal intensity correlates directly with the gene expression level, providing quantitative and sensitive measurement of NFκB target gene activation across multiple samples.