| Cat.No.: | EKIT-0420 |
| Product Name: | Human NFkB p50 Transcription Factor Assay Kit |
| Assay Type: | Semi-quantitative |
| Description: | Human NFkB p50 Transcription Factor Assay Kit is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts and whole cell lysates.A 96-well enzyme-linked immunosorbent assay (ELISA) replaces the cumbersome radioactive electrophoretic mobility shift assay (EMSA). A specific double stranded DNA (dsDNA) sequence containing the NFkB response element is immobilized onto the bottom of wells of a 96-well plate. NFkB contained in a nuclear extract, binds specifically to the NFkB response element. NFkB (p50) is detected by addition of specific primary antibody directed against NFkB (p50). A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm. The kit detects Human NFkB (p50). It will not cross-react with NFkB (p65). |
| Background: | NF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and RelB-p50 complexes are transcriptional activators. The NF-kappa-B p50-p50 homodimer is a transcriptional repressor, but can act as a transcriptional activator when associated with BCL3. NFKB1 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p105 and generation of p50 by a cotranslational processing. The proteasome-mediated process ensures the production of both p50 and p105 and preserves their independent function, although processing of NFKB1/p105 also appears to occur post-translationally. p50 binds to the kappa-B consensus sequence 5'-GGRNNYYCC-3', located in the enhancer region of genes involved in immune response and acute phase reactions. In a complex with MAP3K8, NFKB1/p105 represses MAP3K8-induced MAPK signaling; active MAP3K8 is released by proteasome-dependent degradation of NFKB1/p105. |
| Detection Method: | Colorimetric |
| Target Species: | Human |
| Warning: | For research use only, not for diagnostic use. |
| Product Types | ||
| ◆ Bioactive Small Molecules | ||
| BSM-0182 | NSC 146109 hydrochloride | Inquiry |
| BSM-0404 | Tenovin-3 | Inquiry |
| ◆ Research Kits | ||
| EKIT-0273 | p53 Translocation Assay Kit (Cell-Based) | Inquiry |
| EKIT-0274 | p53 Transcription Factor Activity Assay Kit | Inquiry |
| EKIT-0420 | Human NFkB p50 Transcription Factor Assay Kit | Inquiry |
| Related Gene / Proteins | |||
| p50 | p53 | ||
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