NFkB p65 (pSer536) antibody
Rabbit anti phosphorylated NH-κB p65 (Ser536) antibody (AHP1342) used for the evaluation of NF-κB p65 (pSer536) expression on murine hypopharyngeal primary cells (MHPC) by immunofluorescence.
Image caption:
Pre- or post-application of BAY 11-7082 inhibits the acidic bile-induced nuclear translocation of phospho-NF-κB in MHPC.
Immunofluorescence staining for phospho-NF-κB (p-p65 S536) reveals that 5, 10, or 15 min either of pre- or post-application of BAY 11-7082 inhibits acidic bile-induced p-NF-κB nuclear translocation, demonstrating decreased p-NF-κB nuclear staining and significantly reduced p-NF-κB nuclear expression (p-p65/DAPI ratios of intensity; mean±SD; bin count), compared to MHPC exposed to acidic bile alone. Pre-application of NF-κB inhibitor induces a more intense effect than post-application. Fifteen minutes of pre-application is found to be significantly more effective than its post-application, demonstrating significantly lower nuclear p-NF-κB (p-p65/DAPI) intensity, while no significant differences of the nuclear p-NF-κB (p-p65/DAPI) intensity are observed between 5 or 10 min of pre- and post-application. Control treated-group present a weak p-NF-κB staining [green: p-p65 (S536); blue: DAPI for nuclear staining; p values by t test; multiple comparisons by Holm-Sidak; GraphPad Prism 7.0; p-p65/DAPI ratios of intensity evaluated by Zen imaging software; Zeiss Microscopy].
From: Doukas PG, Vageli DP, Doukas SG, Sasaki CT.
Temporal characteristics of NF-κB inhibition in blocking bile-induced oncogenic molecular events in hypopharyngeal cells.
Oncotarget. 2019 May 21;10(36):3339-51.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.
Rabbit anti NFκB p65 (pSer536) antibody (AHP1342) used to evaluate activated NF-κB expression in murine hypopharyngeal mucosal tissues by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
In vivo topical application of curcumin inhibits acidic bile‐induced activation of NF‐κB. A, IHC analysis for p‐NF‐κB (p65 S536) (from left to right): control untreated HM, showing cytoplasmic staining; saline‐DMSO–treated HM, incorporating sporadic and weak cytoplasmic or nuclear staining in a few basal cells; acidic bile‐treated HM, inducing intense nuclear and cytoplasmic staining of basal and parabasal/suprabasal layers; Pre‐Cur: pre‐application of curcumin + acidic bile‐treated HM, showing weak nuclear or cytoplasmic staining of cells of basal or suprabasal layers; Co‐Cur: co‐application of curcumin + acidic bile‐treated HM, demonstrating cytoplasmic staining of cells of basal layer and weak nuclear or cytoplasmic staining of sporadic cells of suprabasal layers; and Post‐Cur: acidic bile‐treated HM + post‐application of curcumin, demonstrating nuclear or cytoplasmic staining mainly of cells of basal layer and a weak nuclear or cytoplasmic staining of few cells in suprabasal layers.
From: Doukas SG, Doukas PG, Sasaki CT, Vageli D.
The in vivo preventive and therapeutic properties of curcumin in bile reflux-related oncogenesis of the hypopharynx.
J Cell Mol Med. 2020 Sep;24(18):10311-21.
doi: 10.1111/jcmm.15640.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rabbit anti NFkB p65 (pSer536) antibody (AHP1342) used to evaluate NF-κB levels in murine hypopharyngeal primary cells (MHPC) lysates by Western blotting. NB: note incorrect citation of p65 S529 in image caption, should read S536.
Image caption:
Pre- or post-application of ΒΑΥ 11-7082 inhibits acidic bile-induced NF-κB activation and bcl-2 overexpression in MHPC.
Western blot analysis is performed in nuclear and cytoplasmic protein extracts of MHPC for p-NF-κB (p65 S529), and cytoplasmic bcl-2. MHPC exposed to acidic bile demonstrate a significant overexpression of nuclear p-p65 and cytoplasmic bcl-2, compared to controls. Pre- or post-application of BAY 11-7082 results in significantly (A-a) reduced nuclear p-p65 levels compared to acidic bile alone. Pre-application also results in significantly reduced (A-b) cytoplasmic p-p65 levels compared to acidic bile alone. However, post-application of BAY 11-7082 results in elevated cytoplasmic p-p65 levels compared to acidic bile alone. (B) Pre- or post-application of BAY 11-7082 results in significant reduction of cytoplasmic bcl-2 levels compared to acidic bile alone (Paired t-test, *p <0.05; **p <0.005; ***p <0.0005; GraphPad Prism 7.0). (Mean ± SD of three independent experiments). (β-actin and Histone 1 are used for the normalization of cytoplasmic and nuclear protein extracts, respectively).
From: Doukas PG, Vageli DP, Doukas SG, Sasaki CT.
Temporal characteristics of NF-κB inhibition in blocking bile-induced oncogenic molecular events in hypopharyngeal cells.
Oncotarget. 2019 May 21;10(36):3339-3351.
doi: 10.18632/oncotarget.26917.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rabbit anti NFkB p65 (pSer536) antibody (AHP1342) used to stain human hypopharyngeal cells by immunofluorescence.
Image caption:
Silencing of STAT3 suppressed total STAT3, p- STAT3 (Tyr705) and bcl2 protein levels, with a minimal suppressive effect on p- NF- κB (p65 S536), in BA- treated HCs. (A) (a) Immunofluorescence staining for p- STAT3 (Tyr705) (green: p- STAT3; blue: nuclear DNA staining with DAPI; scale bar 20 μm; Zen imagining software). (b) Immunofluorescence staining for p-NF-κB (p65 S536) (green: p-NF-κB (p65 S536); blue: nuclear DNA staining with DAPI; scale bar 20μm; Zen imagining software).
From: Vageli DP, Doukas PG, Siametis A, Judson BL.
Targeting STAT3 prevents bile reflux-induced oncogenic molecular events linked to hypopharyngeal carcinogenesis.
J Cell Mol Med. 2021 Dec 1 [Epub ahead of print].
doi: 10.1111/jcmm.17011.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rabbit anti NFkB p65 (pSer536) antibody (AHP1342) used to stain human hypopharyngeal cells by immunofluorescence.
Image caption:
Pharmacologic inhibition of STAT3 prevents BA- induced nuclear localization of p-STAT3 and Bcl- 2 overexpression, with a minimal effect on p-NF-κB activation, in treated HCs. (B) (a) Immunofluorescence staining for p-NF-κB (p65 S536) (green: p-NF-κB (p65 S536); blue: nuclear DNA staining with DAPI; scale bar 20μm; Zen imagining software).
From: Vageli DP, Doukas PG, Siametis A, Judson BL.
Targeting STAT3 prevents bile reflux-induced oncogenic molecular events linked to hypopharyngeal carcinogenesis.
J Cell Mol Med. 2021 Dec 1 [Epub ahead of print].
doi: 10.1111/jcmm.17011.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rabbit anti NFkB p65 (pSer536) antibody (AHP1342) used to stain human hypopharyngeal cells by immunofluorescence.
Image caption:
Pharmacologic inhibition of STAT3 prevents BA- induced nuclear localization of p-STAT3 and Bcl- 2 overexpression, with a minimal effect on p-NF-κB activation, in treated HCs. (B) (a) Immunofluorescence staining for p-NF-κB (p65 S536) (green: p-NF-κB (p65 S536); blue: nuclear DNA staining with DAPI; scale bar 20μm; Zen imagining software).
From: Vageli DP, Doukas PG, Siametis A, Judson BL.
Targeting STAT3 prevents bile reflux-induced oncogenic molecular events linked to hypopharyngeal carcinogenesis.
J Cell Mol Med. 2021 Dec 1 [Epub ahead of print].
doi: 10.1111/jcmm.17011.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Filter by Application:
IF P WB ResetRabbit anti NFkB p65 (pSer536)
- Product Type
- Polyclonal Antibody
- Isotype
- Polyclonal IgG
- Specificity
- NFkB p65 (pSer536)
Antibody Quality and Performance Guarantee
| Rabbit anti NFkB p65 (pSer536) antibody recognizes nuclear factor NF-kappa-B p65 subunit (NFkB p65), also known as nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 and transcription factor p65, when phosphorylated at serine 536. NFkB is a heterodimeric DNA binding protein which was originally identified consisting of p65 and p50 (NFkB1) subunits. Other subunits, including p52 (NFkB2), cRel, and RelB exist. NFkB is ubiquitously expressed, and functions as a second messenger which upon activation leads to the transcription of a number of genes. NFkB is activated by a number of agents, including pro-inflammatory cytokines and bacterial lipopolysaccharide, and is essential for the maturation of T cells and regulation of their survival and activation. Phosphorylation of serine 536 contributes to NFkB transactivation and increases p65 transcription activity. |
- Target Species
- Human
- Species Cross-Reactivity
-
Target Species Cross Reactivity Rat Expected from Sequence Mouse Expected from Sequence - N.B. Antibody reactivity and working conditions may vary between species.
- Product Form
- Purified IgG - liquid
- Antiserum Preparation
- Antisera to NFkB p65 (pSer536) were raised by repeated immunisation of rabbits with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- <0.1% Sodium Azide (NaN3)
- Immunogen
- Synthetic peptide corresponding to residues surrounding phosphoserine 536 of human NFkB p65 protein.
- Approx. Protein Concentrations
- IgG concentration 0.5mg/ml
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
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 Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |  |
1/500 | 1/3000 |
| Immunohistology - Paraffin | |
1/50 | 1/250 |
| Western Blotting | |
1/100 | 1/1000 |
Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
- Histology Positive Control Tissue
- Human kidney distal tubules and collecting ducts.
- Western Blotting
- AHP1342 detects a band of approximately 65kDa in the appropriate cell lysate.
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References for NFkB p65 (pSer536) antibody
-
Doukas, P.G. et al. (2019) Temporal characteristics of NF-κB inhibition in blocking bile-induced oncogenic molecular events in hypopharyngeal cells.
Oncotarget. 10 (36): 3339-51. -
Doukas, S.G. et al. (2020) The in vivo. preventive and therapeutic properties of curcumin in bile reflux-related oncogenesis of the hypopharynx.
J Cell Mol Med. 24 (18): 10311-21. -
Vageli, D.P. et al. (2021) Targeting STAT3 prevents bile reflux-induced oncogenic molecular events linked to hypopharyngeal carcinogenesis.
J Cell Mol Med. Dec 01 [Epub ahead of print].
Further Reading
-
Mattioli, I. et al. (2004) Transient and selective NF-kappa B p65 serine 536 phosphorylation induced by T cell costimulation is mediated by I kappa B kinase beta and controls the kinetics of p65 nuclear import.
J Immunol. 172 (10): 6336-44.
- RRID
- AB_906096
- UniProt
- Q04206
- Entrez Gene
- RELA
- GO Terms
- GO:0002755 MyD88-dependent toll-like receptor signaling pathway
- GO:0002756 MyD88-independent toll-like receptor signaling pathway
- GO:0003682 chromatin binding
- GO:0003705 sequence-specific enhancer binding RNA polymerase II transcription factor activity
- GO:0005829 cytosol
- GO:0005667 transcription factor complex
- GO:0006916 anti-apoptosis
- GO:0006954 inflammatory response
- GO:0006968 cellular defense response
- View More GO Terms
- GO:0008063 Toll signaling pathway
- GO:0042802 identical protein binding
- GO:0010224 response to UV-B
- GO:0010843 promoter binding
- GO:0016563 transcription activator activity
- GO:0016564 transcription repressor activity
- GO:0019221 cytokine-mediated signaling pathway
- GO:0019901 protein kinase binding
- GO:0031293 membrane protein intracellular domain proteolysis
- GO:0031625 ubiquitin protein ligase binding
- GO:0032582 negative regulation of gene-specific transcription
- GO:0033613 activating transcription factor binding
- GO:0034130 toll-like receptor 1 signaling pathway
- GO:0034134 toll-like receptor 2 signaling pathway
- GO:0034138 toll-like receptor 3 signaling pathway
- GO:0034142 toll-like receptor 4 signaling pathway
- GO:0042301 phosphate binding
- GO:0043123 positive regulation of I-kappaB kinase/NF-kappaB cascade
- GO:0044419 interspecies interaction between organisms
- GO:0045087 innate immune response
- GO:0045944 positive regulation of transcription from RNA polymerase II promoter
- GO:0047485 protein N-terminus binding
- GO:0048011 nerve growth factor receptor signaling pathway
- GO:0050727 regulation of inflammatory response
- GO:0050852 T cell receptor signaling pathway
- GO:0051059 NF-kappaB binding
- GO:0051092 positive regulation of NF-kappaB transcription factor activity
- GO:0051607 defense response to virus
- GO:0070431 nucleotide-binding oligomerization domain containing 2 signaling pathway
- GO:0070491 repressing transcription factor binding
- GO:0070555 response to interleukin-1
AHP1342
148510
180604
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