
Description:
NRF2 plays a crucial role in cellular anti-oxidant defense, making it a therapeutic target for neurodegenerative diseases and cancer. Under unstressed conditions, NRF2 is kept in the cytoplasm by a cluster of proteins that degrade it quickly. Under oxidative stress, NRF2 is not degraded, but instead translocates to the nucleus where it binds to a DNA promoter and initiates gene expression. In the nucleus, NRF2 forms a heterodimer with a small Maf protein and binds to the Antioxidant Response Element in the upstream promoter region of many antioxidative genes, and initiates their transcription. Signosis has developed the NRF2 ELISA kit for the analysis of the NRF2 pathway and to facilitate studying activation of different NRF2-related pathways.Principle:
TF ELISA kit is high sensitive and specific assay with a simple and optimized procedure. The 96-well (8X12 strip) white plate is pre-immobilized with the TF consensus sequencing oligo. The activated TF in nuclear extract or the whole cell lysate is added in the well and binds to the oligo. The activated TF is detected with a specific antibody against the subunit and a HRP conjugated secondary antibody. After incubation, the wells are washed to remove unbound-labeled antibodies. The plate is further detected with HRP luminescent substrate. Luminescence is reported as relative light units (RLUs) on a microplate luminometer. The level of expression for each specific cytokine is directly proportional to the luminescent intensity.
Data:
Analysis of NRF2 activation with NRF2 ELISA in TBHQ-Treated HeLa Cells.
The HeLa cells were treated with 50uM TBHQ overnight. The nuclear extracts were prepared and subjected to NRF2 ELISA kit.