Anti-NFkB p65 antibody - ChIP Grade (ab7970)

Overview

• Product nameAnti-NFkB p65 antibody - ChIP GradeSee all NFkB p65 primary antibodies ...
• Description
  Rabbit polyclonal to NFkB p65 - ChIP Grade
• SpecificityReacts with NFkB p65
• Tested applicationsChIP, ICC, ChIP/Chip, EMSA, IP, IHC-P, WB, IHC-Fr, ICC/IF, Flow Cyt more details
• Species reactivity
  Reacts with: Mouse, Rat, Human
  
• Immunogen

  Synthetic peptide: EDFSSIADMDFSALLSQISS, corresponding to C terminal amino acids 532-551 of Human NFkB p65.

  Run BLAST with Run BLAST with
• Positive controlK-562, NIH/3T3, Hela and A431 cell lysates human DLBCL and human inflammatory skin biopsy
• General notesMembers of the rel/NFkappaB family of transcription factors are involved in the regulation of cellular responses, such as growth, development, and the inflammatory response. They share a structural motif known as the rel homology region (RHR), the C-terminal one third of which mediates protein dimerization. Complexes of p50 (NF-kB1) or p52 (NF-kB2) are generated through the processing of p105 and p100 precursors, respectively. These are usually associated with members of the Rel family (p65, c-Rel, Rel B). The homo- and heterodimer formed through combinations of NF-kB/Rel proteins bind distinct kB sites to regulate the transcription of different genes.

Properties

• FormLiquid
• Storage instructionsStore at +4°C. Do not freeze.
• Storage bufferPBS with 0.1% sodium azide
• Concentration information loading...
• PurityIgG fraction
• Clonality Polyclonal
• IsotypeIgG
• Research Areas
  • Metabolism
  • Types of disease
  • Obesity

  • Cancer
  • Signal transduction
  • Nuclear signaling
  • NFkB pathway

  • Signal Transduction
  • Signaling Pathway
  • Nuclear Signaling
  • NFkB Pathway

  • Cardiovascular
  • Atherosclerosis
  • Vascular Inflammation
  • Inflammatory mediators

  • Microbiology
  • Interspecies Interaction
  • Host Immune Response

  • Microbiology
  • Interspecies Interaction
  • Host Virus Interaction

  • Epigenetics and Nuclear Signaling
  • Nuclear Signaling Pathways
  • NFkB pathway

  • Epigenetics and Nuclear Signaling
  • ChIP'ing antibodies
  • ChIP'ing antibodies

  • Neuroscience
  • Neurology process
  • Neurodegenerative disease
  • Alzheimer's disease
  • Other

  • Cell Biology
  • Apoptosis
  • Intracellular
  • NFkB
  • RelA (p65)

Applications

Target

• FunctionNF-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 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. 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.
• Sequence similaritiesContains 1 RHD (Rel-like) domain.
• Domainthe 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
• Post-translational
  modificationsUbiquitinated, 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.
  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.
  Reversibly acetylated; the acetylation seems to be mediated by CBP, the deacetylation by HDAC3. 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 can also lower DNA-binding and results in nuclear export. Interaction with BRMS1 promotes deacetylation of 'Lys-310'.
• Cellular localizationNucleus. 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.

Target information above from: UniProt accession Q04206 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 5970 Human
  • Entrez Gene: 19697 Mouse
  • Entrez Gene: 309165 Rat
  • Omim: 164014 Human
  • SwissProt: Q04206 Human
  • SwissProt: Q04207 Mouse
  • Unigene: 502875 Human
  • Unigene: 249966 Mouse

  • Unigene: 19480 Rat

  see all

• Alternative names
  Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibodyMGC131774 antibodyNF kappa B p65delta3 antibody

  NFKB 3 antibodyNFKB3 antibodyNuclear Factor NF Kappa B p65 Subunit antibodyNuclear factor NF-kappa-B p65 subunit antibodyNuclear factor of kappa light polypeptide gene enhancer in B cells 3 antibodyNuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B Cells antibodyNuclear factor of kappa light polypeptide gene enhancer in B-cells 3 antibodyOTTHUMP00000233473 antibodyOTTHUMP00000233474 antibodyOTTHUMP00000233475 antibodyOTTHUMP00000233476 antibodyOTTHUMP00000233900 antibodyp65 antibodyp65 NF kappaB antibodyp65 NFkB antibodyRELA antibodyTF65_HUMAN antibodyTranscription Factor p65 antibodyv rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B cells 3 (p65)) antibodyV Rel Avian Reticuloendotheliosis Viral Oncogene Homolog A antibodyv rel reticuloendotheliosis viral oncogene homolog A (avian) antibodyV rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B cells 3, p65 antibodyv-rel reticuloendotheliosis viral oncogene homolog A antibody

  see all

Anti-NFkB p65 antibody - ChIP Grade images

• 

  Western blot - NFkB p65 antibody (ab7970)
  All lanes : Anti-NFkB p65 antibody - ChIP Grade (ab7970)
  
  Lane 1 : A431 whole cell lysate
  Lane 2 : Hela cell lysate
  
  
  Performed under reducing conditions.
  
  Predicted band size : 69 kDa
  Observed band size : 64 kDa (why is the actual band size different from the predicted?)
  
• 

  Immunohistochemistry (Formalin-fixed paraffin-embedded sections) - NFkB p65 antibody (ab7970)
  Immunoperoxidase staining of formalin-fixed, paraffin-embedded human DLBCL showing nuclear expression of NFkB p65 in the tumor cells.
• 

  Immunohistochemistry (Frozen sections) - NFkB p65 antibody - ChIP Grade (ab7970)This image is courtesy of an anonymous Abreview
  ab7970 staining NFkB p65 in Human stomach tissue sections by IHC-Fr (Immunohistochemistry - Froxen sections). Tissue samples were fixed with acetone and blocked with 5% serum for 1 hour at 25°C. Samples were incubated with primary antibody 1/1000 in blocking buffer for 1 hour at 25°C. An undiluted HRP-conjugated Goat polyclonal to rabbit IgG was used as secondary antibody.

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• 

  ChIP - NFkB p65 antibody - ChIP Grade (ab7970)This image was kindly supplied by Mr HoKeun Kwon by Abreview
  ab7970 used undiluted for ChIP analysis of mouse primary T cell nuclear lysate, incubated for 16 hours at 4°C. Cross-linking (X-ChIP) using formaldehyde for 10 minutes.
  
  Detection step: Real-time PCR.
  Negative control: Rabbit IgG.
  
  Results showed the dramatic enrichment of p65 binding upon P+I stimulation in IL-4 promoter region and also IL-2 promoter region.

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• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - NFkB p65 antibody - ChIP Grade (ab7970)This image is a courtesy of Anonymous Abreview

  ab7970 staining NFkB p65 in mouse lung tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent xylene fixation before heat mediated antigen retrieval in citrate and then blocking with 1.5% serum was performed for 30 minutes at 37°C. The primary antibody was used diluted 1/500 and incubated with sample for 16 hours at 4°C. A Biotin conjugated polyclonal to rabbit IgG was used undiluted as secondary antibody. ABC staining system was used.

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• 

  Immunocytochemistry/ Immunofluorescence - NFkB p65 antibody - ChIP Grade (ab7970)This image is courtesy of an anonymous Abreview
  ab7970 staining NFkB p65 in Human AGS cells by ICC/IF (Immunocytochemistry/immunoflurescence). Cells were fixed with formaldehyde, permeabilzed with 0.025% Trton X-100 in TBS and blocked with 5% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/100) for 1 hour at 25°C. An Alexa Fluor^®568-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

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• 

  Western blot - NFkB p65 antibody - ChIP Grade (ab7970)
  Anti-NFkB p65 antibody - ChIP Grade (ab7970) at 1/1000 dilution + Lysate prepared from mouse lung tissue at 20 µg
  
  Secondary
  Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/20000 dilution
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 69 kDa
  Observed band size : 69 kDa
  
  
  Exposure time : 15 seconds

  This image is a courtesy of Anonymous Abreview

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - NFkB p65 antibody - ChIP Grade (ab7970)
  ICC/IF image of ab7970 stained HEK293 cells. The cells were 100% methanol fixed (5 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 (ab7970, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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.