NFkB p65 Antibody (PrecisionAbTM Antibody)

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NFkB p65 Antibody (PrecisionAb<sup>TM</sup> Antibody) gallery image 1

Western blot analysis of whole cell lysates probed with NFkB p65 antibody followed by detection with HRP conjugated Rabbit anti Sheep IgG (H/L) (1/10,000, 5184-2504) and visualized on the ChemiDoc MP with 23 second exposure. Arrow points to NFkB p65 (molecular weight 65 kDa)

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NFkB p65 Antibody (PrecisionAb<sup>TM</sup> Antibody) gallery image 2

Published customer image:
Sheep anti NFκB p65 antibody used for the detection of NFκB p65 expression in HEK293 cell lysates by western blotting
Image caption:
NF-κB p65 protein expression levels (a) Representative immunoblot membrane; (b) Combined densitometry data of three individual experiments, indicating fold changes in the protein expression of NF-κB p65, normalized to the total proteins transferred onto the membrane. BSA: bovine serum albumin; AGEs: advanced glycation end products;* p < 0.05; ** p < 0.01.

From: Serban, A.I.; Stanca, L.; Geicu, O.I.; Dinischiotu, A.
AGEs-Induced IL-6 Synthesis Precedes RAGE Up-Regulation in HEK 293 Cells: An Alternative Inflammatory Mechanism?
Int. J. Mol. Sci. 2015, 16, 20100-20117.

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NFkB p65 Antibody (PrecisionAb<sup>TM</sup> Antibody) gallery image 3

Published customer image:
Sheep anti NFκB antibody (VPA00015) used for the evaluation of NF&kappaB expression in normal human skin derived fibroblasts by western blotting.
Image caption:
AGEs induced signaling in CCD 1070SK fibroblasts. p-ERK1/2 and NF-κB p65 protein expression levels (a) Representative immunoblot membranes; (b) Combined densitometry data indicating fold changes in the protein expression of p-ERK1/2 (b) and NF-κB p65 (c). Immunoreactive bands were normalized to the total proteins transferred onto the membrane in the corresponding lane. Data are relative to BSA and nonimmune IgG treated cells. BSA: bovine serum albumin; AGEs: advanced glycation end products; * p < 0.05; ** p < 0.01; *** p < 0.001.

From: Serban AI, Stanca L, Geicu OI, Munteanu MC, Dinischiotu A (2016)
RAGE and TGF-β1 Cross-Talk Regulate Extracellular Matrix Turnover and Cytokine Synthesis in AGEs Exposed Fibroblast Cells.
PLoS ONE 11(3): e0152376.

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  • Sheep anti NFkB p65
  • NFkB p65 Antibody with Control Lysate
  • NFkB p65 Antibody with Control Lysate
(Rated 0.0 out of 5 based on 0 customer reviews)
    NFkB p65 antibody recognizes transcription factor p65, also known as nuclear factor NF-kappa-B p65 subunit or nuclear factor of kappa light polypeptide gene enhancer in B-cells 3. The antibody has been extensively validated for western blotting using whole cell lysates.
    • Product Type
      PrecisionAb™ Polyclonal
    • Isotype
      Polyclonal IgG
    • Format
      Purified
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      VPA00015WBdatasheet pdfdatasheet pdf100 µl
      VPA00015
      VPA00015KWBdatasheet pdfdatasheet pdf datasheet pdf10 Westerns
      VPA00015K
      VPA00015KTWBdatasheet pdfdatasheet pdf datasheet pdf2 Westerns
      VPA00015KT
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      References
      Reviews
      -
      • Sheep anti Human NFkB p65 antibody recognizes human full length human p65 (NF-kB), also known as transcription factor p65 or nuclear factor of kappa light polypeptide gene enhancer in B-cells 3. Human NFkB p65 is a 551 amino acid pleiotropic transcription factor containing a single RHD domain. Sheep anti Human NFkB p65 antibody will also detect proteolytic fragments of NFkB p65 containing the DNA binding RHD domain located in the amino-terminal region. Multiple isoforms of NFkB p65 are generated by alternative splicing. Sheep anti Human NFkB p65 antibody binds to isoforms 1 and 2 containing the intact RHD domain, but it is unclear whether it will bind to isoforms 3 and 4 which have partial differing deletions within the RHD domain.

        Sheep anti Human NFkB p65 antibody recognizes human NFkB p65 as a single band of ~65 kDa in HEK293 cell line lysates by western blotting under reducing conditions. It also cross reacts with rodent cell line lysates tested.
      • Target Species
        Human
      • Western Blotting
        Anti NFkB p65 antibody detects a band of approximately 65 kDa in HEK293 cell lysates.
        Anti NFkB p65 antibody detects a band of approximately 65 kDa in HEK293 cell lysates.
        Anti NFkB p65 antibody detects a band of approximately 65 kDa in HEK293 cell lysates.
      • Species Cross-Reactivity
        Target SpeciesCross Reactivity
        Mouseyes
        Ratyes
        N.B. Antibody reactivity and working conditions may vary between species.
      • Product Form
        Purified IgG - liquid
        Purified IgG - liquid
        Purified IgG - liquid
      • Preparation
        Sheep polyclonal antibody purified by affinity chromatography
        Pack Size: 10 Westerns100μl Sheep polyclonal antibody purified by affinity chromatography
        Pack Size: 2 Westerns20μl Sheep polyclonal antibody purified by affinity chromatography
        Pack Size: 10 Westerns100μl Sheep polyclonal antibody purified by affinity chromatography
        Pack Size: 2 Westerns20μl Sheep polyclonal antibody purified by affinity chromatography
      • Buffer Solution
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
      • Preservative Stabilisers
        0.09% Sodium Azide (NaN3)
        0.25% Bovine Serum Albumin
        0.09% Sodium Azide (NaN3)
        0.25% Bovine Serum Albumin
        0.09% Sodium Azide (NaN3)
        0.25% Bovine Serum Albumin
      • Immunogen
        Human NFkB p65, amino acids 12-317, expressed in E. coli
      • Instructions for Use
        Please refer to the PrecisionAb western blotting protocol. For additional information on secondary antibody dilution and exposure time see product web page.
        Please refer to the PrecisionAb western blotting protocol. For additional information on secondary antibody dilution and exposure time see product web page.
        Please refer to the PrecisionAb western blotting protocol. For additional information on secondary antibody dilution and exposure time see product web page.
      • Lysate Composition
        400μg HEK293 lysate lyophilized in RIPA buffer.
        400μg HEK293 lysate lyophilized in RIPA buffer.
      • Lysate Reconstitution
        - If using DDT reconstitute the lyophilized lysate with 190μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 10μl 2M DTT.
        - If using BME reconstitute the lyophilized lysate with 180μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 20μl BME.
        Heat at 95°C for 5 minutes. For 10 well mini gels load 25μl. For other gel and comb formats please refer to the PrecisionAb western blotting protocol.
        - If using DDT reconstitute the lyophilized lysate with 190μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 10μl 2M DTT.
        - If using BME reconstitute the lyophilized lysate with 180μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 20μl BME.
        Heat at 95°C for 5 minutes. For 10 well mini gels load 25μl. For other gel and comb formats please refer to the PrecisionAb western blotting protocol.
      • Storage
        Store undiluted at -20oC, avoiding repeated freeze thaw cycles.
        Antibody: Store undiluted at -20oC, avoiding repeated freeze thaw cycles.

        Lysate: Store lyophilized lysate at -20°C. After reconstitution aliquot and store at -20°C for up to 3 months or at -80°C for longer term storage.
        Antibody: Store undiluted at -20oC, avoiding repeated freeze thaw cycles.

        Lysate: Store lyophilized lysate at -20°C. After reconstitution aliquot and store at -20°C for up to 3 months or at -80°C for longer term storage.
      • Shelf Life
        As supplied, 12 months from date of despatch.
        As supplied, 12 months from date of despatch.
        As supplied, 12 months from date of despatch.
      • Acknowledgements
        PrecisionAb™ is a trademark of Bio-Rad Laboratories.
        PrecisionAb™ is a trademark of Bio-Rad Laboratories.
        PrecisionAb™ is a trademark of Bio-Rad Laboratories.
      • Regulatory
        For research purposes only
      • GO Terms
        nerve growth factor receptor signaling pathway
        T cell receptor signaling pathway
        interspecies interaction between organisms
        transcription factor complex
        toll-like receptor 3 signaling pathway
        negative regulation of gene-specific transcription
        positive regulation of I-kappaB kinase/NF-kappaB cascade
        toll-like receptor 2 signaling pathway
        Toll signaling pathway
        response to interleukin-1
        protein kinase binding
        cytosol
        inflammatory response
        regulation of inflammatory response
        positive regulation of transcription from RNA polymerase II promoter
        response to UV-B
        MyD88-independent toll-like receptor signaling pathway
        repressing transcription factor binding
        identical protein binding
        toll-like receptor 1 signaling pathway
        toll-like receptor 4 signaling pathway
        anti-apoptosis
        protein N-terminus binding
        nucleotide-binding oligomerization domain containing 2 signaling pathway
        cytokine-mediated signaling pathway
        promoter binding
        activating transcription factor binding
        sequence-specific enhancer binding RNA polymerase II transcription factor activity
        MyD88-dependent toll-like receptor signaling pathway
        phosphate binding
        membrane protein intracellular domain proteolysis
        transcription repressor activity
        cellular defense response
        defense response to virus
        transcription activator activity
        positive regulation of NF-kappaB transcription factor activity
        chromatin binding
        NF-kappaB binding
        innate immune response
        ubiquitin protein ligase binding
      • UniProt
      • Entrez Gene
      • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
      • Application NameYesNoMin DilutionMax Dilution
        Western Blotting1/1000
      • Application NameYesNoMin DilutionMax Dilution
        Western Blotting1/1000
      • Application NameYesNoMin DilutionMax Dilution
        Western Blotting1/1000

      • PrecisionAb antibodies have been extensively validated for the western blot application. The antibody has been validated at the suggested dilution. Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Further optimization may be required dependant on sample type.
      • Copyright © 2016 Bio-Rad

      Recommended Secondary Antibody

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Rabbit anti Sheep IgG (H/L):HRP5184-25042 mgC, E, WB
        5184-2504
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Rabbit anti Sheep IgG (H/L):HRP5184-25042 mgC, E, WB
        5184-2504
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Rabbit anti Sheep IgG (H/L):HRP5184-25042 mgC, E, WB
        5184-2504

        Recommended Negative Isotype Control

          Useful Reagents

            Recommended Positive Controls

              Histology Controls

                References

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