Anti-NF-kB p65 (acetyl K310) antibody - ChIP Grade (ab52175)

Overview

  • Product nameAnti-NF-kB p65 (acetyl K310) antibody - ChIP GradeSee all NF-kB p65 primary antibodies ...
  • Description
    Rabbit polyclonal to NF-kB p65 (acetyl K310) - ChIP Grade
  • Specificityab52175 detects endogenous levels of NF-kB p65 only when acetylated at lysine 310.
  • Tested applicationsIHC-P, WB, ELISA, ChIP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human NF-kB p65 (internal sequence) (acetyl K310).

  • Positive control
    • Extracts from HeLa cells treated with TSA (40nM, 24hours),human lung carcinoma tissue

Properties

Applications

Our Abpromise guarantee covers the use of ab52175 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
IHC-P 1/50 - 1/100.
WB 1/500 - 1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 60 kDa).
ELISA 1/40000.
ChIP Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 5 µg/ml.

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
    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 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.
  • Information by UniProt
  • Database links
  • Alternative names
    • Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibody
    • MGC131774 antibody
    • NF kappa B p65delta3 antibody
    • NFKB 3 antibody
    • NFKB3 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 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 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 antibody
    • V rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B cells 3, p65 antibody
    see all

Anti-NF-kB p65 (acetyl K310) antibody - ChIP Grade images

  • ab52175 used undiluted in ChIP.
    Whole tissue lysate prepared from murine sciatic nerves.
    10 seconds cross-linking (X-ChIP) with paraformaldehyde.
    Detection step: Semiquantitative PCR.

    See Abreview

  • ICC/IF image of ab52175 stained HeLa 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 (ab52175, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

  • Immunofluorescence analysis of ab52175 stained HeLa cells.The picture on the right is treated with the synthesized peptide.


  • developed using the ECL technique

    Predicted band size : 60 kDa


    Exposure time : 30 seconds

    Image courtesy of Tom Hu by Abreview.

    ab52175 used at a 1/1000 dilution in Western Blot.Whole tissue lysate prepared from murine sciatic nerves was loaded at 200&microg.Performed under reduced, denaturing conditions.Blocking step 5% milk for 1 hour at 20°C.Secondary used was an HRP conjugated goat anti-rabbit IgG polyclonal used at a 1/5000 dilution.

    See Abreview

  • All lanes : Anti-NF-kB p65 (acetyl K310) antibody - ChIP Grade (ab52175) at 1/500 dilution

    Lane 1 : HeLa cell extract treated with TSA (40nM, 24hours) with immunising peptide
    Lane 2 : HeLa cell extract treated with TSA (40nM, 24hours)

    Secondary
    Alkaline Phosphatase-conjugated Monoclonal Mouse Anti-Rabbit.

    Predicted band size : 60 kDa
    Observed band size : 70 kDa (why is the actual band size different from the predicted?)
  • Anti-NF-kB p65 (acetyl K310) antibody - ChIP Grade (ab52175) + Extracts from HeLa cells,trated with TSA 40nM 24h

    Predicted band size : 60 kDa
  • Immunohistochemistry analysis of paraffin-embedded Human lung carcinoma tissue with ab52175.The picture on the right is treated with synthesized peptide.

References for Anti-NF-kB p65 (acetyl K310) antibody - ChIP Grade (ab52175)

This product has been referenced in:
  • Ziesché E  et al. The coactivator role of histone deacetylase 3 in IL-1-signaling involves deacetylation of p65 NF-?B. Nucleic Acids Res 41:90-109 (2013). Read more (PubMed: 23087373) »
  • Price LC  et al. Nuclear Factor ?-B Is Activated in the Pulmonary Vessels of Patients with End-Stage Idiopathic Pulmonary Arterial Hypertension. PLoS One 8:e75415 (2013). IHC-Fr ; Human . Read more (PubMed: 24124488) »

See all 8 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (8% in MES)
Sample Human Tissue lysate - whole (Superior Frontal Cortex)
Specification Superior Frontal Cortex
Blocking step Li-Cor Block Buffer as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 100% · Temperature: 21°C
Username

Grant Corbett

Verified customer

Submitted Jul 15 2014

Thank you for contacting us and I am sorry for the delay in getting back to you.

I now have some further information to share with you in regards to the anti-NFkB p65 (acetyl K310) antibody (ab52175) and how its been used in ELISA. The protoco...

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Thank you for your reply.

I have arranged a free of charge vial of ab52175 to ship out to Cedarlane on their next order from us as a replacement for the faulty ab19870 that you ordered through them on reference number is 389794. When they get...

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Thank you for taking time to contact us. I am sorry to hear that this antibody is not providing satisfactory results.

Do you know your lot number and order number for this antibody?

Do you have a copy of your WB in electronic form...

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Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guar...

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Application ChIP
Sample Mouse Tissue lysate - whole (sciatic nerves)
Specification sciatic nerves
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: paraformaldehyde
Detection step Semiquantitative PCR
Positive control Control
Negative control HDAC 1/2 cko
Username

Mr. Tom Hu

Verified customer

Submitted Mar 23 2012

Application Western blot
Sample Mouse Tissue lysate - whole (Sciatic nerves)
Loading amount 200 µg
Specification Sciatic nerves
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Mr. Tom Hu

Verified customer

Submitted Mar 23 2012

Thank you for contacting us.

We are excited that you are aware of our Abreview testing program. The program is designed so that researchers may test products in previously untested species or applications without the worry as it entitles the...

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Application Western blot
Sample Mouse Cell lysate - nuclear (C2C12 wild type (RS) and PC4siRNA (RS/2))
Loading amount 30 µg
Specification C2C12 wild type (RS) and PC4siRNA (RS/2)
Treatment GM, 24hDM, 48hDM
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Felice Tirone

Verified customer

Submitted Jun 21 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"