Product: NF-kB p65 Antibody
Catalog: AF6387
Description: Rabbit polyclonal antibody to NF-kB p65
Application: WB IHC IF/ICC IP
Cited expt.: WB, IHC, IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep, Dog, Xenopus
Mol.Wt.: 65kDa; 60kD(Calculated).
Uniprot: Q04206
RRID: AB_2835228

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:200, IP
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(91%), Bovine(100%), Horse(100%), Sheep(100%), Dog(100%), Xenopus(82%)
Clonality:
Polyclonal
Specificity:
NF-kB p65 Antibody detects endogenous levels of total NF-kB p65.
RRID:
AB_2835228
Cite Format: Affinity Biosciences Cat# AF6387, RRID:AB_2835228.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

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;

Immunogens

Immunogen:

A synthesized peptide derived from human NF-kB p65, corresponding to a region within the internal amino acids.

Uniprot:
Gene(ID):
Description:
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.
Sequence:
MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKEDIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELSEPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Zebrafish
91
Xenopus
82
Chicken
70
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

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. 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.

PTMs:

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.

Subcellular Location:

Nucleus. Cytoplasm.
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).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Family&Domains:

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).

Research Fields

· 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.

References

1). The Circadian Clock Gene Bmal1 Regulates Microglial Pyroptosis After Spinal Cord Injury via NF-κB/MMP9. CNS neuroscience & therapeutics, 2024 (PubMed: 39648661) [IF=4.8]

Application: WB    Species: Mouse    Sample: spinal cord

FIGURE 4. Bmal1 can effectively inhibit NF‐κB in vitro and in vivo. (a) The expression of p‐NF‐κB p65 in the spinal cord of mice was detected by Western blot and quantitative statistical analysis (n = 3). (b) The expression of p‐NF‐κB p65 in the spinal cord of mice in each group was detected by immunohistochemistry and statistical analysis (n = 3, scale bar = 50 μm, the relative content of p‐NF‐κB p65 was calculated by Image J software). (c, d) Western blot was used to detect the expression of p‐NF‐κB p65 in BV2 cells after up‐regulation or down‐regulation of Bmal1 and quantitative analysis (n = 3). (e, f) Immunofluorescence was used to detect the expression of p‐NF‐κB p65 in BV2 cells after up‐regulation and down‐regulation of Bmal1 and quantitative statistical analysis (n = 3, scale bar = 50 μm, the relative content of p‐NF‐κB p65 was calculated by Image J software). (All the data are expressed as means ± SD, one‐way ANOVA followed by Tukey's post hoc test was applied ^^^ p 

Application: IHC    Species: Mouse    Sample: spinal cord

FIGURE 4. Bmal1 can effectively inhibit NF‐κB in vitro and in vivo. (a) The expression of p‐NF‐κB p65 in the spinal cord of mice was detected by Western blot and quantitative statistical analysis (n = 3). (b) The expression of p‐NF‐κB p65 in the spinal cord of mice in each group was detected by immunohistochemistry and statistical analysis (n = 3, scale bar = 50 μm, the relative content of p‐NF‐κB p65 was calculated by Image J software). (c, d) Western blot was used to detect the expression of p‐NF‐κB p65 in BV2 cells after up‐regulation or down‐regulation of Bmal1 and quantitative analysis (n = 3). (e, f) Immunofluorescence was used to detect the expression of p‐NF‐κB p65 in BV2 cells after up‐regulation and down‐regulation of Bmal1 and quantitative statistical analysis (n = 3, scale bar = 50 μm, the relative content of p‐NF‐κB p65 was calculated by Image J software). (All the data are expressed as means ± SD, one‐way ANOVA followed by Tukey's post hoc test was applied ^^^ p 

Application: IF/ICC    Species: Mouse    Sample: spinal cord

FIGURE 4. Bmal1 can effectively inhibit NF‐κB in vitro and in vivo. (a) The expression of p‐NF‐κB p65 in the spinal cord of mice was detected by Western blot and quantitative statistical analysis (n = 3). (b) The expression of p‐NF‐κB p65 in the spinal cord of mice in each group was detected by immunohistochemistry and statistical analysis (n = 3, scale bar = 50 μm, the relative content of p‐NF‐κB p65 was calculated by Image J software). (c, d) Western blot was used to detect the expression of p‐NF‐κB p65 in BV2 cells after up‐regulation or down‐regulation of Bmal1 and quantitative analysis (n = 3). (e, f) Immunofluorescence was used to detect the expression of p‐NF‐κB p65 in BV2 cells after up‐regulation and down‐regulation of Bmal1 and quantitative statistical analysis (n = 3, scale bar = 50 μm, the relative content of p‐NF‐κB p65 was calculated by Image J software). (All the data are expressed as means ± SD, one‐way ANOVA followed by Tukey's post hoc test was applied ^^^ p 

2). Recurrent non-severe hypoglycemia aggravates cognitive decline in diabetes and induces mitochondrial dysfunction in cultured astrocytes. Molecular and Cellular Endocrinology, 2021 (PubMed: 33545179) [IF=3.8]

Application: WB    Species: mice    Sample: cortex tissue

Fig. 3. Recurrent non-severe hypoglycemia (RH) inhibits brain-derived neurotrophic factor (BDNF) and glial cell derived neurotrophic factor (GDNF) signaling in diabetic mice. (A) The expression of BDNF/tropomyosin (t)-receptor (r)-kinase (k) B (TrkB) signaling and GDNF/GDNF family receptor alpha-1 (GFRa1)/receptor tyrosine kinase (Ret) signaling in mice cerebral cortex, measured using western blotting. (B–G) Relative protein levels of BDNF, TrkB, p-TrkB, GDNF, GFRa1, Ret, and p-Ret (Tyr1062), as determined by ImageJ. *P < 0.05 vs. normal control (NC), **P < 0.01 vs. NC, #P < 0.05 vs. diabetes mellitus (DM), ##P < 0.05 vs. DM.

3). LncRNA BANCR promotes oral squamous cell carcinoma progression via regulating Rab1A signaling. Journal of Oral Pathology & Medicine, 2023 (PubMed: 37433101) [IF=2.7]

4). Astragaloside IV regulates NF‑κB‑mediated cellular senescence and apoptosis of hepatic stellate cells to suppress PDGF‑BB‑induced activation. Experimental and Therapeutic Medicine, 2019 (PubMed: 31641375) [IF=2.4]

Application: WB    Species: rat    Sample: HSC‑T6

Figure 6. |The expression of components in the NF‑κB pathway in platelet‑derived growth factor‑BB‑activated HSC‑T6 treated with ASIV. (A‑E) The relative mRNA expression levels of (A) p65, (B) p52, (C) p50, (D) IKKα and (E) IκBα. (F) Protein expression of p65, p52 and IκBα. *P<0.05 and **P<0.01. IKKα, inhibitor of nuclear factor‑κB kinase subunit‑α; IκBα, inhibitor of nuclear factor‑κB‑α; NF‑κB, nuclear factor‑κB; ASIV, astragaloside IV.

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