|Product Overview:||NF-κB is a transcription factor that plays a pivotal role in a number of physiological cell processes such as growth, apoptosis, and immune response through regulating the expression of a number of cellular genes. Normally, NF-κB is inactive and retained in the cytoplasm bound to its inhibitory protein IκB. Upon stimulation of various factors such as infection of pathogens including viruses and bacteria or treatment of cytokines such as IL-1 or TNF-α, IκB proteins are phosphorylated, ubiquitinated and finally degraded. Released NF-κB proteins then form homo- or heterodimers, most commonly p50/p65 or p50/p50, which are transported into the nucleus where they bind to specific DNA sequences of targeted genes and activate transcription. NF-κB also induces expression of its own inhibitor, IκBα, which binds to active NF-κB in the nucleus to terminate the induction of gene expression. The NF-κB p65 Transcription Factor-Activity Assay kit is a non-radioactive transcription factor assay with an ELISA format. It offers an easy, speedy, sensitive and high-throughput method to detect the activation of transcription factors. In 96-well plates, double stranded oligonucleotides containing NF-κB binding sequence have been coated. These oligonucleotides specifically capture the active NF-κB p65 contained in whole cell lysate or nuclear extracts after a short incubation. The specificity of the reaction between active NF-κB p65 and the DNA probe is additionally stringent because of the establishment of specific competitive DNA and non-specific competitive DNA probes in this reaction system.|
|Applications:||Detecting the NF-κB p65 in human nuclear extraction and whole lysates.|
|Kit Components:||Microplate; DNA Binding Buffer (5X); Positive Control; Specific Competitor DNA Probe; Non-specific Competitor DNA Probe; Assay Reagent; DTT (300 mM); Wash Buffer Concentrate (20X); Primary Antibody; HRP-conjugated Secondary Antibody; Antibody Diluent Buffer; TMB One-Step Substrate Reagent; Stop Solution|
|Detection method:||Absorbance (450 nm)|
|Features & Benefits:||A non-radioactive transcription factor assay with an ELISA format.
An easy, speedy, sensitive and high-throughput method to detect the activation of transcription factors.
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