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    nf-kb-p65-antibody-ab140751.pdf

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Cell Biology Apoptosis Intracellular NFkB RelA (p65)
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Validated using a knockout cell line

Anti-NF-kB p65 antibody (ab140751)

  • Datasheet
Reviews (2) Submit a question References (2)

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Western blot - Anti-NF-kB p65 antibody (ab140751)
  • Western blot - Anti-NF-kB p65 antibody (ab140751)

Key features and details

  • Chicken polyclonal to NF-kB p65
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Rat, Human
  • Isotype: IgY

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Overview

  • Product name

    Anti-NF-kB p65 antibody
    See all NF-kB p65 primary antibodies
  • Description

    Chicken polyclonal to NF-kB p65
  • Host species

    Chicken
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse, Cow, Pig, Chimpanzee, Macaque monkey, Gorilla, Chinese hamster
  • Immunogen

    Synthetic peptide within Human NF-kB p65 aa 500 to the C-terminus conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.

  • Positive control

    • This antibody gave a positive signal in the following lysates: HeLa whole cell; Jurkat whole cell; Rat spleen tissue.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 3% BSA
  • Concentration information loading...
  • Purity

    Protein L purified
  • Clonality

    Polyclonal
  • Isotype

    IgY
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • NFkB
    • RelA (p65)
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFkB pathway
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • NFkB pathway
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Microbiology
    • Interspecies Interaction
    • Host Immune Response
    • Metabolism
    • Types of disease
    • Obesity
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • NFkB
    • RelA (p65)

Associated products

  • Isotype control

    • Chicken IgY - Isotype Control (ab50579)
  • Recombinant Protein

    • Recombinant Human NF-kB p65 protein (Tagged) (ab114150)
  • Related Products

    • NF kappaB p65 ELISA Kit (ab176648)

Applications

Our Abpromise guarantee covers the use of ab140751 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).

Target

  • Function

    NF-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 similarities

    Contains 1 RHD (Rel-like) domain.
  • Domain

    the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
  • Post-translational
    modifications

    Ubiquitinated, 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 localization

    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.
  • 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: 466665 Chimpanzee
    • Entrez Gene: 508233 Cow
    • Entrez Gene: 101150285 Gorilla
    • Entrez Gene: 5970 Human
    • Entrez Gene: 19697 Mouse
    • Entrez Gene: 100135665 Pig
    • 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 antibody
    • MGC131774 antibody
    • NF kappa B p65delta3 antibody
    • nfkappabp65 antibody
    • NFkB p65 antibody
    • NFKB3 antibody
    • Nuclear factor kappaB antibody
    • Nuclear Factor NF Kappa B p65 Subunit antibody
    • Nuclear factor NF-kappa-B p65 subunit antibody
    • Nuclear factor of kappa light polypeptide gene enhancer in B cells 3 antibody
    • Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 antibody
    • OTTHUMP00000233473 antibody
    • OTTHUMP00000233474 antibody
    • OTTHUMP00000233475 antibody
    • OTTHUMP00000233476 antibody
    • OTTHUMP00000233900 antibody
    • p65 antibody
    • p65 NF kappaB antibody
    • p65 NFkB antibody
    • relA antibody
    • TF65_HUMAN antibody
    • Transcription factor NFKB3 antibody
    • Transcription factor p65 antibody
    • v rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B cells 3 (p65)) antibody
    • V rel avian reticuloendotheliosis viral oncogene homolog A antibody
    • v rel reticuloendotheliosis viral oncogene homolog A (avian) antibody
    • V rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B cells 3, p65 antibody
    see all

Images

  • Western blot - Anti-NF-kB p65 antibody (ab140751)
    Western blot - Anti-NF-kB p65 antibody (ab140751)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: NF-κB p60 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: A431 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab140751 observed at 70 kDa. Red - ab8245 loading control, observed at 37 kDa.


    ab140751 was shown to specifically react with NF-κB p65 in wild-type HAP1 cells. No band was observed when NF-κB p65 knockout samples were used. Wild-type and NF-κB p65 knockout samples were subjected to SDS-PAGE. ab140751 (NF-κB p65) and ab8245 (loading control to GAPDH) were diluted 1 μg/mL and 1/2000 and incubated overnight at 4°C. Blots were developed with IRDye® 800CW Donkey anti-Chicken IgG (H + L) and Donkey Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216779 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-NF-kB p65 antibody (ab140751)
    Western blot - Anti-NF-kB p65 antibody (ab140751)
    All lanes : Anti-NF-kB p65 antibody (ab140751) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : Spleen (Rat) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Chicken IgY H&L (HRP) (ab6877) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 65 kDa
    Observed band size: 65 kDa
    Additional bands at: 120 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute

Protocols

  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (2)

    Publishing research using ab140751? Please let us know so that we can cite the reference in this datasheet.

    ab140751 has been referenced in 2 publications.

    • Wang Y  et al. Sevoflurane alleviates LPS-induced acute lung injury via the microRNA-27a-3p/TLR4/MyD88/NF-?B signaling pathway. Int J Mol Med 44:479-490 (2019). PubMed: 31173183
    • Wang H  et al. Low-protein diets supplemented with casein hydrolysate favor the microbiota and enhance the mucosal humoral immunity in the colon of pigs. J Anim Sci Biotechnol 10:79 (2019). PubMed: 31624591

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunocytochemistry abreview for Anti-NF-kB p65 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (Para influenza virus infected lung epithelial cell)
    Permeabilization
    Yes - Triton X100 (0.1%)
    Specification
    Para influenza virus infected lung epithelial cell
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Selvarani Vimalanathan

    Verified customer

    Submitted May 09 2016

    Western blot abreview for Anti-NF-kB p65 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Non-reduced Denaturing (8%)
    Sample
    Human Cell lysate - whole cell (MEFs, U2OS, HEK 293 and Fibroblasts)
    Specification
    MEFs, U2OS, HEK 293 and Fibroblasts
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 05 2014

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