Product nameAnti-NFkB p100 (phospho S865) antibody
See all NFkB p100 primary antibodies
DescriptionRabbit polyclonal to NFkB p100 (phospho S865)
SpecificityThis antibody detects endogenous levels of NFkB p100 protein only when phosphorylated at Serine 865.
Tested applicationsSuitable for: ICC/IF, WB, IHC-P, ELISA, IPmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
Synthetic peptide within Human NFkB p100 (phospho S865). The exact sequence is proprietary.
- WB: extracts of ovary cancer cells. IHC-P:breast carcinoma tissue.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride
Without Mg2+ and Ca2+
Concentration information loading...
PurityImmunogen affinity purified
Purification notesThis antibody was affinity purified from rabbit antiserum by affinity chromatography using epitope specific phosphopeptide.
Our Abpromise guarantee covers the use of ab31474 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||1/500 - 1/1000. Predicted molecular weight: 54 kDa.|
|IHC-P||1/50 - 1/100.|
|IP||Use at an assay dependent concentration.|
RelevanceNF-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. 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. In a non-canonical activation pathway, the MAP3K14-activated CHUK/IKKA homodimer phosphorylates NFKB2/p100 associated with RelB, inducing its proteolytic processing to NFKB2/p52 and the formation of NF-kappa-B RelB-p52 complexes. The NF-kappa-B heterodimeric RelB-p52 complex is a transcriptional activator. The NF-kappa-B p52-p52 homodimer is a transcriptional repressor. NFKB2 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p100 and generation of p52 by a cotranslational processing. The proteasome-mediated process ensures the production of both p52 and p100 and preserves their independent function. p52 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. p52 and p100 are respectively the minor and major form; the processing of p100 being relatively poor. Isoform p49 is a subunit of the NF-kappa-B protein complex, which stimulates the HIV enhancer in synergy with p65. In concert with RELB, regulates the circadian clock by repressing the transcriptional activator activity of the CLOCK-ARNTL/BMAL1 heterodimer.
Cellular localizationCytoplasmic and Nuclear
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Ab31474, at a dilution of 1/50, staining NFkB p100 in paraffin embedded human breast carcinoma tissue by Immunohistochemistry.
Left image: treated with ab31474.
Right image: treated with the same antibody preincubated with sythesized peptide.
All lanes : Anti-NFkB p100 (phospho S865) antibody (ab31474) at 1/500 dilution
Lane 1 : Extracts of ovary cancer cells (5-30ug).
Lane 2 : Extracts of ovary cancer cells (5-30ug). Antibody pre-incubated with synthesized peptide.
All lanes : goat-anti-rabbit IgG-AP-conjugate.
Predicted band size: 54 kDa
ICC/IF image of ab31474 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab31474, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab31474 has not yet been referenced specifically in any publications.