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    nf-kb-p65-acetyl-k310-antibody-ab19870.pdf

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Cell Biology Apoptosis Intracellular NFkB RelA (p65)
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Anti-NF-kB p65 (acetyl K310) antibody (ab19870)

  • Datasheet
Reviews (6)Q&A (4)References (92)

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Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
  • Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
  • Dot Blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
  • Immunoprecipitation - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
  • Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
  • Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)

Key features and details

  • Rabbit polyclonal to NF-kB p65 (acetyl K310)
  • Suitable for: WB, IP, Dot blot
  • Reacts with: Mouse, Rat
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-NF-kB p65 (acetyl K310) antibody
    See all NF-kB p65 primary antibodies
  • Description

    Rabbit polyclonal to NF-kB p65 (acetyl K310)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, Dot blotmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat
    Predicted to work with: Human
  • Immunogen

    Synthetic peptide corresponding to Human NF-kB p65 aa 300-400 (internal sequence) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab20612)

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • 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

  • Assay kits

    • NFkB p65 Transcription Factor Assay Kit (ab133112)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

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

    • Human NF-kB p65 peptide (ab16636)
    • NF kappaB p65 ELISA Kit (ab176648)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab19870 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 (3)
Use a concentration of 2.5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa). Collaborator data suggests that immunoprecipitation of this antibody prior to Western blotting is required to obtain the best results (see images)
IP
Use a concentration of 2.5 µg/ml.
Dot blot
Use at an assay dependent concentration.
Notes
WB
Use a concentration of 2.5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa). Collaborator data suggests that immunoprecipitation of this antibody prior to Western blotting is required to obtain the best results (see images)
IP
Use a concentration of 2.5 µg/ml.
Dot blot
Use at an assay dependent concentration.
Application notes
Is unsuitable for ICC/IF.

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: 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
    • 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 (acetyl K310) antibody (ab19870)
      Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)This image is courtesy of Prof. Michael Hottiger's Lab, University of Zurich
      Lanes 1-2 : Coomassie stain
      Lanes 3-4 : Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 1/500 dilution

      Lanes 1 & 3 : Acetylated p65 protein
      Lanes 2 & 4 : K310R mutant protein

      Lysates/proteins at 2 µg per lane.

      Secondary
      Lanes 3-4 : IRDye® 800CW Goat anti-Rabbit IgG (H + L) at 1/15000 dilution

      Predicted band size: 65 kDa



      Observed band size: 75kDa

      The p65 band runs higher in this blot because the protein contains a myc-tag.

    • Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
      Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)This image is courtesy of an Abreview submitted by Christian Marx
      All lanes : Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 1/1000 dilution

      Lane 1 : HCT116 whole cell lysate treated with DMSO for 24 hrs (control)
      Lane 2 : HCT116 whole cell lysate treated with 2 µM SAHA for 24 hrs
      Lane 3 : MEF whole cell lysate treated with DMSO for 24 hrs (control)
      Lane 4 : MEF whole cell lysate treated with 2 µM SAHA for 24 hrs

      Lysates/proteins at 60 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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: 140 kDa (possible non-specific binding), 15 kDa (possible non-specific binding), 40 kDa (possible non-specific binding), 45 kDa (possible non-specific binding), 90 kDa (possible non-specific binding)


      Exposure time: 2 minutes

      See Abreview

    • Dot Blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
      Dot Blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)This image is courtesy of Christine Buerki and Karin Rothgiesser, University of Zurich, Switzerland

      Rabbit polyclonal to NF-kB p65 (acetyl K310) (ab19870; 2.5µg/ml) in 1% non-fat milk TBS-T incubated for 3h at room temperature. Exposure time: 75 min normal ECL. This Dot blot demonstrates that ab19870 recognized upto 10ng of purified peptide on a PVDF membrane.

    • Immunoprecipitation - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
      Immunoprecipitation - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)This image is courtesy of Christine Buerki and Karin Rothgiesser, University of Zurich, Switzerland

      Western Blot with ab19870 after p65 Immunoprecipitation: rabbit polyclonal to NF-kB p65 (acetyl K310) (ab19870; 2.5µg/ml) in 1% non-fat milk TBS-T incubated for 3 hours at room temperature. Exposure time: 1 min normal ECL. Tested samples: nuclear extracts (180 µg) of immortalized p65-/- mouse cells, complemented with the empty vector (pRRL), wild-type p65 (Wt) and non-acetylatable K310 (K310R). The samples tested were treated with deacetylase inhibitors HDACi (TSA + Nicotinamide) and TNF-alpha. The samples were immunoprecipitated with 2µg of alpha-p65 and subsequently analysed by Western blot with Rabbit polyclonal to NF-kB p65 (acetyl K310) (ab19870). Predicted band size = 65kDa, Observed band size = 75kDa. The p65 band runs higher in this SDS-PAGE blot as it contains a myc-tag. 

    • Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
      Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)This image is courtesy of Christine Buerki and Karin Rothgiesser, University of Zurich, Switzerland
      All lanes : Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 2.5 µg/ml

      Lane 1 : pRRL untreated
      Lane 2 : pRRL HDACi
      Lane 3 : pRRL HDACi + TNF
      Lane 4 : Wt untreated
      Lane 5 : Wt HDACi
      Lane 6 : Wt HDACi + phorbol myristate acetate
      Lane 7 : K310R untreated
      Lane 8 : K310R HDACi
      Lane 9 : K310R HDACi + phorbol myristate acetate

      Lysates/proteins at 75 µg per lane.

      Developed using the ECL technique.

      Predicted band size: 65 kDa
      Observed band size: 75 kDa why is the actual band size different from the predicted?


      Exposure time: 1 hour


      ab19870 recognizes Rabbit polyclonal to NF-kB p65 (acetyl K310) specifically at ~75kDa (indicated by the arrow) is this SDS-PAGE blot. The p65 band runs higher than 65kDa in this SDS-PAGE blot as it contains a myc-tag. We are sure that the band at ~75kDa is p65 since p65 specific antibodies detect the same band in IP and WB and there is no signal in the p65 knock-out cell line with ab19870. A number of additonal bands are recognized by ab19870 when tested with endogenous p65 from whole cell extracts, we do not know the identity of these bands.

      Tested samples: nuclear extracts (75µg) of immortalized p65-/- mouse cells, complemented with the empty ve

    • Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
      Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
      Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 1 µg/ml + Lung (Rat) Tissue Lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Performed under reducing conditions.

      Predicted band size: 65 kDa
      Observed band size: 72 kDa why is the actual band size different from the predicted?
      Additional bands at: 15 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 4 minutes

    Protocols

    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (92)

    Publishing research using ab19870? Please let us know so that we can cite the reference in this datasheet.

    ab19870 has been referenced in 92 publications.

    • Ma Q  et al. MicroRNA-210 downregulates TET2 and contributes to inflammatory response in neonatal hypoxic-ischemic brain injury. J Neuroinflammation 18:6 (2021). PubMed: 33402183
    • Lee SU  et al. Transforming Growth Factor ß Inhibits MUC5AC Expression by Smad3/HDAC2 Complex Formation and NF-?B Deacetylation at K310 in NCI-H292 Cells. Mol Cells 44:38-49 (2021). PubMed: 33510050
    • Chen L  et al. Mangiferin prevents myocardial infarction-induced apoptosis and heart failure in mice by activating the Sirt1/FoxO3a pathway. J Cell Mol Med 25:2944-2955 (2021). PubMed: 33523605
    • Zhou B  et al. Bortezomib suppresses self-renewal and leukemogenesis of leukemia stem cell by NF-?B-dependent inhibition of CDK6 in MLL-rearranged myeloid leukemia. J Cell Mol Med 25:3124-3135 (2021). PubMed: 33599085
    • Cheng Y  et al. iTRAQ-Based Quantitative Proteomics Indicated Nrf2/OPTN-Mediated Mitophagy Inhibits NLRP3 Inflammasome Activation after Intracerebral Hemorrhage. Oxid Med Cell Longev 2021:6630281 (2021). PubMed: 33628368
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-NF-kB p65 (acetyl K310) antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Placenta)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    15 µg
    Specification
    Placenta
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Jeremiah Gaudet

    Verified customer

    Submitted Aug 23 2018

    Western blot abreview for Anti-NFkB p65 (acetyl K310) antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    100000 cells
    Gel Running Conditions
    Reduced Denaturing (12 %)
    Sample
    Human Cell lysate - whole cell (breast)
    Specification
    breast
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 20 2013

    Western blot abreview for Anti-NFkB p65 (acetyl K310) antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HCT116 colon cancer cell line and MEF cell line)
    Loading amount
    60 µg
    Specification
    HCT116 colon cancer cell line and MEF cell line
    Treatment
    DMSO control and 2 µM SAHA for 24 hrs
    Gel Running Conditions
    Reduced Denaturing (13%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Mr. Christian Marx

    Verified customer

    Submitted Feb 11 2013

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