|Application ||WB, IHC|
|Reactivity||Human, Mouse, Rat|
|Calculated MW||60212 Da|
|Application & Usage||Western blotting (0.5-4 µg/ml), immunocytochemistry (10-20 µg/ml). However, the optimal conditions should be determined individually. The antibody detects primarily NFkB p65 in samples of human, mouse, and rat origins.|
|Other Names||RELA, NFKB3, MGC131774|
|Formulation||100 µg (0.2 mg/ml) affinity purified rabbit anti-NFkB polyclonal antibody in phosphate buffered saline (PBS), pH 7.2, containing 25% glycerol, 0.5% BSA, 0.01% thimerosal.|
|Handling||The antibody solution should be gently mixed before use.|
|Reconstitution & Storage||-20 °C|
|Precautions||NFkB p65 Antibody is for research use only and not for use in diagnostic or therapeutic procedures.|
|Function||NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes 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 p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression (By similarity). The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA- binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells (By similarity).|
|Cellular Location||Nucleus. Cytoplasm. Note=Colocalized with DDX1 in the nucleus upon TNF-alpha induction (By similarity) Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalizes with GFI1 in the nucleus after lipopolysaccharide (LPS) stimulation|
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Provided below are standard protocols that you may find useful for product applications.
NFkB is a heterodimer that consists of a 50 kDa DNA binding subunit (p50) and a 65 kDa transactivation subunit (p65/RelA). Both of these subunits exhibit sequence homology to the protooncogene c-Rel. The p50 has an isoform called p49/p52, and both proteins are derived from the amino-terminal of precursor protein p105 and p100. The p50/p65 heterodimer remains in the cytosol in an inactive form as a complex with its inhibitor, IkB. Upon stimulation of cells by a wide variety of stimuli such as lipopolysaccharide (LPS), pro-inflammatory cytokines (IL-1 & TNF, etc.), and viral infection, IkB is phosphorylated and degraded by proteosome. The active NFkB heterodimer is translocated into the nucleus and induces gene expression.
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