NF-kB (p65) Transcription Factor Assay Kit is a non-radioactive method for detecting specific transcription factor DNA binding activity.
The NF-κB/Rel family of transcription factors is comprised of several structurally-related proteins that form homodimers and heterodimers and include p50/p105, p52/p100, RelA (p65), c-Rel/NF-κB. Members of this family are responsible for regulating over 150 target genes, including the expression of inflammatory cytokines, chemokines, immunoreceptors, and cell adhesion molecules. Because of this, NF-κB has often been called a"central mediator of the human immune response. Acting as dimers, these transcription factors bind to DNA sequences, collectively called κB, sites thereby regulating expression of target genes. In most cells, Rel/ NF-κB transcription complexes are present in an inactive form in the cytoplasm, bound to an inhibitor IκB. Certain stimuli result in the phosphorylattion, ubiquitination and subsequent degradation of IκB proteins thereby enabling translocation of NF-κB into the nucleus. The most common Rel/NF-κB dimer in mammals contains p50-RelA (p50/ p65) heterodimers and is specifically called NF-κB. One of the target genes activated by NF-κB is that encoding IκBα. This feedback mechanism allows newly-synthesized IκBα to enter the nucleus, remove NF-κB from DNA and transport it back to the cytoplasm thereby restoring its inactive state. The importance of Rel/NF-κB transcription factors in human inflammation and certain diseases makes them attractive targets for potential therapeutics.
For research use only (RUO)
Kit components may be stored at -20°C prior to use. For long term storage, the Positive Control should be thawed on ice, aliquoted at 20 μL/vial, and stored at -80°C. After use we recommend each kit component be stored according to the temperature listed below.This kit will perform as specified if stored as directed and used before the expiration date indicated on the outside of the box.Item StorageTranscription Factor Binding Assay Buffer (4X) 4°CTranscription Factor Reagent A -20°CTranscription Factor NF-kB (human p65) Positive Control -80°CTranscription Factor Antibody Binding Buffer (10X) 4°CTranscription Factor NF-kB (p65) Primary Antibody -20°CWash Buffer Concentrate (400X) RTPolysorbate 20 RTTranscription Factor NF-kB Competitor dsDNA -20°CTranscription Factor Goat Anti-Rabbit HRP Conjugate -20°CTranscription Factor NF-kB 96-Well Strip Plate 4°C96-Well Cover Sheet RTTranscription Factor Developing Solution 4°CTranscription Factor Stop Solution RT
Transcription Factor Binding Assay Buffer (4X) 1 vialTranscription Factor Reagent A 1 vialTranscription Factor NF-kB (human p65) Positive Control 1 vialTranscription Factor Antibody Binding Buffer (10X) 1 vialTranscription Factor NF-kB (p65) Primary Antibody 1 vialWash Buffer Concentrate (400X) 1 vialPolysorbate 20 1 vialTranscription Factor NF-kB Competitor dsDNA 1 vialTranscription Factor Goat Anti-Rabbit HRP Conjugate 1 vialTranscription Factor NF-kB 96-Well Strip Plate 1 plate96-Well Cover Sheet 1 coverTranscription Factor Developing Solution 1 vialTranscription Factor Stop Solution 1 vial