Avian reticuloendotheliosis viral (v rel) oncogene homolog A; MGC131774; NF kappa B p65delta3; NFKB3; Nuclear Factor NF Kappa B p65 Subunit; Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B cells 3; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3; OTTHUMP00000233473; OTTHUMP00000233474; OTTHUMP00000233475; OTTHUMP00000233476; OTTHUMP00000233900; p65; p65 NF kappaB; p65 NFkB; relA; TF65_HUMAN; Transcription factor p65; v rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B cells 3 (p65)); V rel avian reticuloendotheliosis viral oncogene homolog A; v rel reticuloendotheliosis viral oncogene homolog A (avian); V rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B cells 3, p65;
WB 1:500-1:2000, IHC 1:50-1:500, IP 1:100-1:500, IF/ICC 1:200, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
Human, Mouse, Rat, Monkey
Pig, Bovine, Horse, Sheep, Dog
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Phospho-NF-kB p65 (Ser536) Antibody detects endogenous levels of NF-kB p65 only when phosphorylated at Serine 536.
Please cite this product as: Affinity Biosciences Cat# AF2006, RRID:AB_2834435.
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, sodium azide and glycerol.Store at -20 °C.Stable for 12 months from date of receipt.
A synthesized peptide derived from human NF- kappaB p65 around the phosphorylation site of Ser536.
Observed Mol.Wt.: 65 kD.
Predicted Mol.Wt.: 60kDa(Calculated)..
Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalized with RELA in the nucleus upon TNF-alpha induction.
NFKB1 (MIM 164011) or NFKB2 (MIM 164012) is bound to REL (MIM 164910), RELA, or RELB (MIM 604758) to form the NFKB complex. The p50 (NFKB1)/p65 (RELA) heterodimer is the most abundant form of NFKB. The NFKB complex is inhibited by I-kappa-B proteins (NFKBIA, MIM 164008 or NFKBIB, MIM 604495), which inactivate NFKB by trapping it in the cytoplasm.
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. The heterodimeric RELA-NFKB1 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. The NF-kappa-B heterodimeric RELA-NFKB1 and RELA-REL complexes, for instance, function as transcriptional activators. 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. The inhibitory effect of I-kappa-B on NF-kappa-B through retention in the cytoplasm is exerted primarily through the interaction with RELA. RELA shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Beside its activity as a direct transcriptional activator, it is also able to modulate promoters accessibility to transcription factors and thereby indirectly regulate gene expression. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells. The NF-kappa-B homodimeric RELA-RELA complex appears to be involved in invasin-mediated activation of IL-8 expression.
Ubiquitinated by RNF182, leading to its proteasomal degradation. Degradation is required for termination of NF-kappa-B response.
Monomethylated at Lys-310 by SETD6. Monomethylation at Lys-310 is recognized by the ANK repeats of EHMT1 and promotes the formation of repressed chromatin at target genes, leading to down-regulation of NF-kappa-B transcription factor activity. Phosphorylation at Ser-311 disrupts the interaction with EHMT1 without preventing monomethylation at Lys-310 and relieves the repression of target genes (By similarity).
Phosphorylation at Ser-311 disrupts the interaction with EHMT1 and promotes transcription factor activity (By similarity). Phosphorylation on Ser-536 stimulates acetylation on Lys-310 and interaction with CBP; the phosphorylated and acetylated forms show enhanced transcriptional activity. Phosphorylation at Ser-276 by RPS6KA4 and RPS6KA5 promotes its transactivation and transcriptional activities.
Reversibly acetylated; the acetylation seems to be mediated by CBP, the deacetylation by HDAC3 and SIRT2. Acetylation at Lys-122 enhances DNA binding and impairs association with NFKBIA. Acetylation at Lys-310 is required for full transcriptional activity in the absence of effects on DNA binding and NFKBIA association. Acetylation at Lys-310 promotes interaction with BRD4. Acetylation can also lower DNA-binding and results in nuclear export. Interaction with BRMS1 promotes deacetylation of Lys-310. Lys-310 is deacetylated by SIRT2.
S-nitrosylation of Cys-38 inactivates the enzyme activity.
Sulfhydration at Cys-38 mediates the anti-apoptotic activity by promoting the interaction with RPS3 and activating the transcription factor activity.
Sumoylation by PIAS3 negatively regulates DNA-bound activated NF-kappa-B.
Proteolytically cleaved within a conserved N-terminus region required for base-specific contact with DNA in a CPEN1-mediated manner, and hence inhibits NF-kappa-B transcriptional activity.
Note: Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B) (PubMed:1493333). Colocalized with DDX1 in the nucleus upon TNF-alpha induction (PubMed:19058135). Colocalizes with GFI1 in the nucleus after LPS stimulation (PubMed:20547752). Translocation to the nucleus is impaired in L.monocytogenes infection (PubMed:20855622).
Component of the NF-kappa-B p65-p50 complex. Component of the NF-kappa-B p65-c-Rel complex. Homodimer; component of the NF-kappa-B p65-p65 complex. Component of the NF-kappa-B p65-p52 complex. May interact with ETHE1. Binds TLE5 and TLE1. Interacts with TP53BP2. Binds to and is phosphorylated by the activated form of either RPS6KA4 or RPS6KA5. Interacts with ING4 and this interaction may be indirect. Interacts with CARM1, USP48 and UNC5CL. Interacts with IRAK1BP1 (By similarity). Interacts with NFKBID (By similarity). Interacts with NFKBIA. Interacts with GSK3B. Interacts with NFKBIB (By similarity). Interacts with NFKBIE. Interacts with NFKBIZ. Interacts with EHMT1 (via ANK repeats) (By similarity). Part of a 70-90 kDa complex at least consisting of CHUK, IKBKB, NFKBIA, RELA, ELP1 and MAP3K14. Interacts with HDAC3; HDAC3 mediates the deacetylation of RELA. Interacts with HDAC1; the interaction requires non-phosphorylated RELA. Interacts with CBP; the interaction requires phosphorylated RELA. Interacts (phosphorylated at 'Thr-254') with PIN1; the interaction inhibits p65 binding to NFKBIA. Interacts with SOCS1. Interacts with UXT. Interacts with MTDH and PHF11. Interacts with ARRB2. Interacts with NFKBIA (when phosphorylated), the interaction is direct; phosphorylated NFKBIA is part of a SCF(BTRC)-like complex lacking CUL1. Interacts with RNF25. Interacts (via C-terminus) with DDX1. Interacts with UFL1 and COMMD1. Interacts with BRMS1; this promotes deacetylation of 'Lys-310'. Interacts with NOTCH2 (By similarity). Directly interacts with MEN1; this interaction represses NFKB-mediated transactivation. Interacts with AKIP1, which promotes the phosphorylation and nuclear retention of RELA. Interacts (via the RHD) with GFI1; the interaction, after bacterial lipopolysaccharide (LPS) stimulation, inhibits the transcriptional activity by interfering with the DNA-binding activity to target gene promoter DNA. Interacts (when acetylated at Lys-310) with BRD4; leading to activation of the NF-kappa-B pathway. Interacts with MEFV. Interacts with CLOCK (By similarity). Interacts (via N-terminus) with CPEN1; this interaction induces proteolytic cleavage of p65/RELA subunit and inhibition of NF-kappa-B transcriptional activity. Interacts with FOXP3. Interacts with CDK5RAP3; stimulates the interaction of RELA with HDAC1, HDAC2 and HDAC3 thereby inhibiting NF-kappa-B transcriptional activity. Interacts with DHX9; this interaction is direct and activates NF-kappa-B-mediated transcription. Interacts with LRRC25. Interacts with TBX21 (By similarity). Interacts with KAT2A (By similarity). Interacts with ZBTB7A; involved in the control by RELA of the accessibility of target gene promoters. Directly interacts with DDX3X; this interaction may trap RELA in the cytoplasm, impairing nuclear relocalization upon TNF activating signals.
(Microbial infection) Interacts with human respiratory syncytial virus (HRSV) protein M2-1.
(Microbial infection) Interacts with molluscum contagiosum virus MC132.
(Microbial infection) Interacts with herpes virus 8 virus protein LANA1.
The transcriptional activation domain 3/TA3 does not participate to the direct transcriptional activity of RELA but is involved in the control by RELA of the accessibility of target gene promoters. Mediates interaction with ZBTB7A.
The transcriptional activation domain 1/TA1 and the transcriptional activation domain 2/TA2 have direct transcriptional activation properties (By similarity). The 9aaTAD motif found within the transcriptional activation domain 2 is a conserved motif present in a large number of transcription factors that is required for their transcriptional transactivation activity (PubMed:17467953).
· Cellular Processes > Cell growth and death > Apoptosis.(View pathway)
· Cellular Processes > Cell growth and death > Cellular senescence.(View pathway)
· Environmental Information Processing > Signal transduction > MAPK signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Ras signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > cAMP signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > TNF signaling pathway.(View pathway)
· Human Diseases > Drug resistance: Antineoplastic > Antifolate resistance.
· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.
· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).
· Human Diseases > Substance dependence > Cocaine addiction.
· Human Diseases > Infectious diseases: Bacterial > Epithelial cell signaling in Helicobacter pylori infection.
· Human Diseases > Infectious diseases: Bacterial > Shigellosis.
· Human Diseases > Infectious diseases: Bacterial > Salmonella infection.
· Human Diseases > Infectious diseases: Bacterial > Pertussis.
· Human Diseases > Infectious diseases: Bacterial > Legionellosis.
· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.
· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).
· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.
· Human Diseases > Infectious diseases: Parasitic > Amoebiasis.
· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.
· Human Diseases > Infectious diseases: Viral > Hepatitis C.
· Human Diseases > Infectious diseases: Viral > Hepatitis B.
· Human Diseases > Infectious diseases: Viral > Measles.
· Human Diseases > Infectious diseases: Viral > Influenza A.
· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.
· Human Diseases > Infectious diseases: Viral > HTLV-I infection.
· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.
· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.
· Human Diseases > Cancers: Overview > Viral carcinogenesis.
· Human Diseases > Cancers: Specific types > Pancreatic cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Prostate cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia.(View pathway)
· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.(View pathway)
· Human Diseases > Cancers: Specific types > Small cell lung cancer.(View pathway)
· Human Diseases > Immune diseases > Inflammatory bowel disease (IBD).
· Organismal Systems > Immune system > Chemokine signaling pathway.(View pathway)
· Organismal Systems > Aging > Longevity regulating pathway.(View pathway)
· Organismal Systems > Development > Osteoclast differentiation.(View pathway)
· Organismal Systems > Immune system > Toll-like receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > NOD-like receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > RIG-I-like receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > Cytosolic DNA-sensing pathway.(View pathway)
· Organismal Systems > Immune system > IL-17 signaling pathway.(View pathway)
· Organismal Systems > Immune system > Th1 and Th2 cell differentiation.(View pathway)
· Organismal Systems > Immune system > Th17 cell differentiation.(View pathway)
· Organismal Systems > Immune system > T cell receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > B cell receptor signaling pathway.(View pathway)
· Organismal Systems > Nervous system > Neurotrophin signaling pathway.(View pathway)
· Organismal Systems > Endocrine system > Prolactin signaling pathway.(View pathway)
· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.
· Organismal Systems > Endocrine system > Relaxin signaling pathway.
Application: WB Species:mouse; Sample:Not available
Figure 7. FCN-A promoted the M1 polarization of BMDMs through a TLR4/MyD88-dependent pathway in vitro. (a) The protein expressions of the purified GST-FCN-A and GST were determined by SDS-PAGE. (b) The extracted membrane proteins from RAW264.7 cells were incubated with the purified GST-FCN-A or GST proteins. Co-IP analysis of the interaction between TLR4 of macrophage and GST-FCN-A was performed by using anti-TLR4. Rabbit IgG was used as a negative control in co-IP. (c, e) BMDMs, isolated from WT, TLR4-/- or MyD88-/- mice, were stimulated with FCN-A (10g/mL) for 24 h, then the expressions of iNOS and Arg-1 from BMDMs were examined by Western blot analysis. (d, f) The levels of pro-inflammatory cytokines IL-1in cell lysates, and secreted IL-6, TNF- were detected by ELISA. (g, h) Western blot analysis of p-IRAK1, p-p65, p-ERK1/2, and p-JNK in the BMDM lysates of TLR4-/-, MyD88-/- or WT after stimulation with FCN-A for 0-45 min. In d and f, values are mean ± [SEM] from three independent experiments.
Application: WB Species:rat; Sample:Not available
FIGURE 3: Pro-inflammatory factors, anti-oxidant enzyme and cell signaling pathways in resveratrol (Res)- or vehicle-treated cystic kidneys. (A) TNF-α, MCP-1, CFB and SOD2 were analyzed by western blot in 9-week-old +/+ and Cy/+ kidneys. (B–E) Immunohistochemical staining for oxidative stress markers 8-OHdG and nitrotyrosine in the tubulointerstitial area. Computer-assisted morphometry was used to quantify changes of 8-OHdG and nitrotyrosine in each group. Scale bar = 50 µm. (F) NF-κB pathway ( p-p65, p65, p105 and p50) and mTOR pathway ( p-S6K and total S6K) were analyzed by western blot in 9-week-old +/+ and Cy/+ kidneys. Blots are representative of three independent experiments.
Application: WB Species:mouse; Sample:Not available
(c) Western blotting analyses of total NF-κ B p65 and phospho-NF-κB p65 in primary murine chondrocytes with or without IL-1β (10 ng/ml) for 24h. Increased phospho-NF-κB p65 protein expression in chondrocytes from AMPKα cDKO mice compared with their WT littermates was observed. GAPDH served as a loading control.
Application: IHC Species:mouse; Sample:Not available
Figure 6. IHC analyses of surgically induced and ageing-associated OA. (a,c and e) Representative IHC images of MMP-3, MMP-13 and phospho-NF-κB p65 in the medial tibial plateau in AMPKα1α2 conditional double knockout (AMPKα cDKO) mice and their WT littermates 2 weeks post-sham operation and DMM surgery or in mice at 9 months of age. Scale bars=20μm. The cellularity of the section was confirmed with haematoxylin staining.
Application: WB Species:human; Sample:human
Figure 3. Survivin overexpression results in upregulation of NF-κBp65, IKKα, IΚΚβ in Eca109 and KYSE150cells. (A and B) Eca109 and (C and D) KYSE150 cells were transfected with GV142-survivin overexpression plasmid (Eca109 survivin OE and KYSE150 survivin OE), GV142-control plasmid (Eca109 control and KYSE150 control), and mock transfection. Expression levels of survivin, NF-κB p65, IΚΚβ, and IΚΚα were analyzed by (A and C) RT‑qPCR and (B and D) western blotting of total protein. Transcript levels were measured by RT-qPCR of total isolated RNA, with GADPH serving as an internal control. Columns indicate the mean values from triplicate experiments and the error bars indicate standard deviation. * P<0.05; **P
Application: WB Species:human; Sample:Not available
Fig 5. The expression of p-p65, p-AKT and Bcl-2 (protein level) in U2-OS cells treated by LY294002 or/and Estrogen receptor β (ERβ) siRNA in the presence of 10-10 M E2.
Tips: For phospho antibody, we provide phospho peptide（0.5mg) and non-phospho peptide(0.5mg).
Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.
Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.
This product is for research use only. Not for use in diagnostic or therapeutic procedures.
|S276||Phosphorylation||P11309 (PIM1) , O94806 (PRKD3) , O75676 (RPS6KA4) , P17612 (PRKACA) , O75582 (RPS6KA5)||Uniprot|
|S468||Phosphorylation||P49841 (GSK3B) , O14920 (IKBKB) , Q14164 (IKBKE)||Uniprot|
|S529||Phosphorylation||P68400 (CSNK2A1) , P47710 (CSN1S1)||Uniprot|
|S536||Phosphorylation||O15111 (CHUK) , O94806 (PRKD3) , Q9Y6K9 (IKBKG) , Q9UHD2 (TBK1) , Q9HCP0 (CSNK1G1) , Q16566 (CAMK4) , P51812 (RPS6KA3) , Q15418 (RPS6KA1) , Q00534 (CDK6) , O14920 (IKBKB) , P24723 (PRKCH) , Q14164 (IKBKE)||Uniprot|