Phospho-NF-κB p65 (Ser536) Recombinant antibody

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  WB analysis using 80379-2-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 80379-2-RR (Phospho-NF-κB p65 (Ser536) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of NIH/3T3 using 80379-2-RR

  Non-treated and Calyculin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 80379-2-RR (Phospho-NF-κB p65 (Ser536) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Actin (66009-1-Ig) antibody as a loading control.

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  FC experiment of PC-3 using 80379-2-RR

  1x10^6 untreated or Calyculin A treated PC-3 cells were intracellularly stained with 0.25 ug Phospho-NF-κB p65 (Ser536) Recombinant antibody (80379-2-RR, Clone:240777D9) and PE-Conjugated Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue), 1x10^6 untreated PC-3 cells were intracellularly stained with 0.25 ug Phospho-NF-κB p65 (Ser536) Recombinant antibody (80379-2-RR, Clone:240777D9) and PE-Conjugated Goat Anti-Rabbit IgG(H+L)(black). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

Phospho-NF-κB p65 (Ser536) Uni-rAb^TM Recombinant Antibody for WB, FC (Intra), ELISA

Cat No. 80379-2-RR

Clone No.240777D9

Publications(7)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (1)

Applications

WB, FC (Intra), ELISA

Synonyms

NF-κB p65 (Ser536), NF-κB p65 Ser-536, Phospho NF-κB p65 (Ser536), phospho-NF-κB p65, Phospho-NF-κB p65 Ser536

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Tested Applications

• Positive WB detected in: Calyculin A treated NIH/3T3 cells, Calyculin A treated HeLa cells
• Positive FC (Intra) detected in: PC-3 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 7 publications below

Product Information

80379-2-RR targets Phospho-NF-κB p65 (Ser536) in WB, FC (Intra), ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Peptide
• Full Name: v-rel reticuloendotheliosis viral oncogene homolog A (avian)
• Calculated Molecular Weight: 65 kDa
• Observed Molecular Weight: 75 kDa
• GenBank Accession Number: BC011603
• Gene Symbol: NF-κB p65
• Gene ID (NCBI): 5970
• RRID: AB_3670476
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• UNIPROT ID: Q04206
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Nuclear factor kB (NF-kB) is a collective term for a small family of dimeric transcription factors [comprising p65 (RelA) and RelB, c-Rel, p50/p105 (NF-κB1), and p52/p100 (NF-κB2)]. All NF-κB proteins share a Rel homology domain (RHD), which is responsible for DNA binding and dimerization. Only p65, RelB, and c-Rel contain potent transactivation domains within sequences from the C-terminal to the RHD. Exterior signals lead to the phosphorylation and degradation of the inhibitory complex IκB, which is modulated by the IκB kinase (IKK), and its degradation allows for the release of the typical NF-κB heterodimer, p65/p50, to translocate into the nucleus. NF-κB binds to its cognate DNA elements and can transcriptionally activate different target genes among which 200-500 genes have been implicated in cell survival/apoptosis, cell growth, immune response, and inflammation.

Publications

Species	Application	Title
mouse	WB	Int J Nanomedicine; Unexpected Inhibitory Role of Silica Nanoparticles on Lung Cancer Development by Promoting M1 Polarization of Macrophages; Authors - Meng Xiang
human	WB	Front Immunol; SQLE-mediated squalene metabolism promotes tumor immune evasion in pancreatic cancer; Authors - Junchen Pan
mouse,rat	WB	Cell Death Discov; 17β-estradiol alleviated ferroptotic neuroinflammation by suppressing ATF4 in mouse model of Parkinson's disease; Authors - Guoming Wang
mouse	WB	Adv Mater; Suppression of Sepsis Cytokine Storm by Escherichia Coli Cell Wall-Derived Carbon Dots; Authors - Yinan Li
mouse	WB	Front Vet Sci; Overexpression of the Mas1 gene mitigated LPS-induced inflammatory injury in mammary epithelial cells by inhibiting the NF-κB/MAPKs signaling pathways; Authors - Shuping Yan
human	WB	Microbiol Spectr; Mutation of Pseudomonas aeruginosa lasI/rhlI diminishes its cytotoxicity, oxidative stress, inflammation, and apoptosis on THP-1 macrophages; Authors - Yanying Ren