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AB239882

Anti-NF-kB p65 (phospho S536) 抗体 [EP2294Y] - BSA and Azide free

Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free

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(56 Publications)

Rabbit Recombinant Monoclonal NF-kB p65 phospho S536 antibody. Carrier free. Suitable for IP, Dot, WB and reacts with Human, Mouse, Rat, Synthetic peptide samples. Cited in 56 publications.

別名を表示する

NFKB3, RELA, Transcription factor p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3

5 Images
Immunoprecipitation - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)
  • IP

Unknown

Immunoprecipitation - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

ab76302 at 1/30 immunoprecipitating NF-kB p65 (phospho S536) in Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.

Lane 1 (input) : Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha (10μg)

Lane 2 (+) : ab76302 + Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab76302 in Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.

For western blotting, ab76302 at 1/500 dilution (0.95 μg/ml) and VeriBlot for IP Detection Reagent (HRP)(ab131366) at 1/1000 dilution were used.

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76302).

All lanes:

Immunoprecipitation - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (<a href='/products/primary-antibodies/nf-kb-p65-phospho-s536-antibody-ep2294y-ab76302'>ab76302</a>)

Predicted band size: 60 kDa

Observed band size: 65 kDa

false

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)
  • WB

Lab

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

This data was developed using ab76302, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000,000 dilution.

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 100ng/ml Calyculin A for 10 minutes (Lane 2).

All lanes:

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (<a href='/products/primary-antibodies/nf-kb-p65-phospho-s536-antibody-ep2294y-ab76302'>ab76302</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa whole cell lysate at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate treated with 100ng/ml Calyculin A for 10 minutes at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 65 kDa,36 kDa

false

Exposure time: 5s

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)
  • WB

Unknown

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76302).

All lanes:

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (<a href='/products/primary-antibodies/nf-kb-p65-phospho-s536-antibody-ep2294y-ab76302'>ab76302</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes whole cell lysate at 15 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 60 kDa

Observed band size: 65 kDa

false

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)
  • WB

Unknown

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

All lanes:

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (<a href='/products/primary-antibodies/nf-kb-p65-phospho-s536-antibody-ep2294y-ab76302'>ab76302</a>) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg

Lane 2:

C6 (Rat glial tumor glial cell) treated with 100nM Calyculin A for 30 minutes whole cell lysate at 15 µg

Lane 3:

C6 (Rat glial tumor glial cell) treated with 100nM Calyculin A for 30 minutes, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 60 kDa

Observed band size: 65 kDa

false

Dot Blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)
  • Dot

Unknown

Dot Blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

Dot blot analysis of INF- kB p65 (phospho S536) phospho peptide (Lane 1) and NF- kB p65 non-phospho peptide (Lane 2) labeling NF-kB p65 (phospho S536) with ab76302 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) at 1/100 000 was used as the secondary antibody.

Blocking and diluting buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76302).

関連する標識済み抗体及び組成の異なる製品 (1)

  • Unconjugated

    Anti-NF-kB p65 (phospho S536) antibody [EP2294Y]

Key facts

宿主種

Rabbit

クローン性

Monoclonal

クローン番号

EP2294Y

アイソタイプ

IgG

キャリアフリー

Yes

交差種

Mouse, Rat, Human

アプリケーション

WB, IP, Dot

applications

免疫原

The exact immunogen used to generate this antibody is proprietary information.

特異性

<p>Stimulation may be required to allow detection of the phosphorylated protein.<br>We recommend using NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes as a positive control.</p>

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代替製品

Primary Antibodies

AB190589

Alexa Fluor® 647 Anti-NF-kB p65 antibody [E379]

RabMAb|Recombinant

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-NF-kB p65 antibody [E379] (AB190589)

  • 4
  • 3
Primary Antibodies

AB76311

Anti-NF-kB p65 antibody [EP2161Y]

20ul selling size|RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

  • 4
  • 1
Primary Antibodies

AB207297

Anti-NF-kB p65 antibody [E379] - BSA and Azide free

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 antibody [E379] - BSA and Azide free (AB207297)

  • 3
  • 1
Primary Antibodies

AB32536

Anti-NF-kB p65 antibody [E379]

BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NF-kB p65 antibody [E379] (AB32536)

  • 4
  • 15
Primary Antibodies

AB190205

Alexa Fluor® 488 Anti-NF-kB p65 antibody [E379]

RabMAb|Recombinant

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-NF-kB p65 antibody [E379] (AB190205)

  • 0
  • 0
Primary Antibodies

AB109458

Anti-NF-kB p65 (phospho S529) antibody [EPR2379]

20ul selling size|RabMAb|Recombinant

Western blot - Anti-NF-kB p65 (phospho S529) antibody [EPR2379] (AB109458)

  • 0
  • 0

組合せ製品

Assay Kits

AB176663

NF kappaB p65 (pS536 + Total) ELISA Kit

SimpleStep

ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)

  • 0
  • 0
Sample Prep & Detection Kits

AB269822

Alexa Fluor® 594 Conjugation Kit (Fast) - Lightning-Link®

Schematic Diagram - Alexa Fluor® 594 Conjugation Kit (Fast) - Lightning-Link® (AB269822)

  • 5
  • 1
Sample Prep & Detection Kits

AB218260

Oligonucleotide Conjugation Kit

Conjugation - Oligonucleotide Conjugation Kit (AB218260)

  • 0
  • 0
Primary Antibodies

AB32536

Anti-NF-kB p65 antibody [E379]

BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NF-kB p65 antibody [E379] (AB32536)

  • 4
  • 15
Primary Antibodies

AB32518

Anti-IKB alpha antibody [E130]

20ul selling size|RabMAb|Recombinant|KO Validated

Western blot - Anti-IKB alpha antibody [E130] (AB32518)

  • 4
  • 31
Primary Antibodies

AB133462

Anti-IKB alpha (phospho S36) antibody [EPR6235(2)]

20ul selling size|RabMAb|Recombinant

Flow Cytometry (Intracellular) - Anti-IKB alpha (phospho S36) antibody [EPR6235(2)] (AB133462)

  • 4
  • 1
Primary Antibodies

AB92700

Anti-IKB alpha (phospho S32) antibody [EPR3148]

20ul selling size|RabMAb|Recombinant

Immunoprecipitation - Anti-IKB alpha (phospho S32) antibody [EPR3148] (AB92700)

  • 5
  • 1
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<p></p>", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>" }, "Mouse": { "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<p></p>", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>" }, "Rat": { "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<p></p>", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>" }, "Synthetic peptide": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "<p></p>", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }
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製品の詳細

ab239882 is the carrier-free version of ab76302.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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出荷温度及び保存条件

製品の状態
Liquid
精製方法
Affinity purification Protein A
バッファー組成
pH: 7.2 - 7.4 Constituents: PBS
出荷温度
Blue Ice
短期保存温度
+4°C
長期保存温度
+4°C
保管に関する情報
Do Not Freeze
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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

NF-kB p65 also known as RelA is a significant component of the NF-kB protein complex. This complex usually involves a molecular weight for p65 of approximately 65 kDa. NF-kB p65 is mechanistically a transcription factor that regulates genes involved in inflammation cell survival and immune response. Expression of p65 is widely seen in various cell types including immune cells and epithelial cells suggesting its role in numerous physiological processes.
Biological function summary

NF-kB p65 acts as part of the larger NF-kB complex usually forming a heterodimer with other family members like p50. This complex translocates to the nucleus upon activation where it binds specific DNA sequences to regulate gene expression. The p65 subunit is essential for transactivating target genes involved in immune and inflammatory responses. Its regulation is important for proper cellular functioning especially in the maintenance of immune homeostasis and inflammatory response.

Pathways

NF-kB p65 participates in critical pathways like the NF-kB signaling and Toll-like receptor pathways. These pathways are fundamental for initiating immune responses and contribute to the regulation of apoptosis and cellular stress responses. In the context of these pathways the IKK complex plays a pivotal role in NF-kB activation leading to the phosphorylation and subsequent degradation of inhibitors that retain NF-kB p65 in the cytoplasm therefore allowing its movement to the nucleus.

NF-kB p65 is implicated in conditions such as cancer and autoimmune diseases. Its dysregulation can lead to persistent activation of inflammatory pathways contributing to tumorigenesis and chronic inflammation. In the context of these diseases proteins such as the p50 subunit and the IKK complex are often involved reflecting the importance of tight regulation of NF-kB signaling in preventing pathological conditions.
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製品プロトコール

For this product, it's our understanding that no specific protocols are required. You can visit:
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ターゲットの情報

NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes 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. The heterodimeric RELA-NFKB1 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. The NF-kappa-B heterodimeric RELA-NFKB1 and RELA-REL complexes, for instance, function as transcriptional activators. 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. The inhibitory effect of I-kappa-B on NF-kappa-B through retention in the cytoplasm is exerted primarily through the interaction with RELA. RELA shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Besides its activity as a direct transcriptional activator, it is also able to modulate promoters accessibility to transcription factors and thereby indirectly regulate gene expression. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells (PubMed : 15790681). The NF-kappa-B homodimeric RELA-RELA complex appears to be involved in invasin-mediated activation of IL-8 expression. Key transcription factor regulating the IFN response during SARS-CoV-2 infection (PubMed : 33440148).
See full target information RELA pS536
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文献 (56)

Recent publications for all applications. Explore the full list and refine your search

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Quercetin as a therapeutic agent activate the Nrf2/Keap1 pathway to alleviate lung ischemia-reperfusion injury.

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Mohammad Yousefi Zardak,Fatemeh Keshavarz,Ali Mahyaei,Morteza Gholami,Fatemeh Sadat Moosavi,Elham Abbasloo,Farzaneh Abdollahi,Maryam Hossein Rezaei,Elham Madadizadeh,Nasrin Soltani,Fatemeh Bejeshk,Niyan Salehi,Fahimeh Rostamabadi,Fatemeh Bagheri,Mahla Jafaraghae,Mahdiyeh Ranjbar Zeydabadi,Meraj Baghgoli,Gholamreza Sepehri,Mohammad Abbas Bejeshk

Frontiers in pharmacology 15:1449985 PubMed39263577

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Protective effects of insulin on dry eye syndrome via TLR4/NF-κB pathway: based on network pharmacology and experiments validation.

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Ruolan Huang,Shenglan Gong,Bocheng Xiong,Xifei Yang,Chongyang Chen,Wei Song,Ruodai Wu,Li Yang,Jia Yin,Mingtai Chen
View all publications
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