NF-κB p65 Recombinant antibody

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  WB analysis using 80979-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 80979-1-RR (NF-κB p65 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 80979-1-RR

  WB result of NF-κB p65 antibody (80979-1-RR; 1:13000; incubated at room temperature for 1.5 hours) with sh-Control and sh-NF-κB p65 transfected HEK-293 cells.

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  WB analysis of HEK-293 using 80979-1-RR

  WB result of NF-κB p65 antibody (80979-1-RR; 1:10000; incubated at room temperature for 1.5 hours) with sh-Control and sh-NF-κB p65 transfected HEK-293 cells.

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  IP experiment of HeLa using 80979-1-RR

  IP result of anti-NF-κB p65 (IP:80979-1-RR, 4ug; Detection:80979-1-RR 1:4000) with HeLa cells lysate 1085 ug.

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  IF Staining of HT-1080 using 80979-1-RR

  Immunofluorescent analysis of (4% PFA) fixed TNF alpha treated HT-1080 cells using NF-κB p65 antibody (80979-1-RR, Clone: 4C7 ) at dilution of 1:400 and CoraLite®594-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-4), CL488-Phalloidin (green).

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  IF Staining of HepG2 using 80979-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using NF-κB p65 antibody (80979-1-RR, Clone: 4C7 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  IF Staining of HepG2 using 80979-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using NF-κB p65 antibody (80979-1-RR, Clone: 4C7 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of HepG2 using 80979-1-RR

  1X10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human NF-κB p65 (80979-1-RR, Clone:4C7) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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  ChIP experiment of HeLa using 80979-1-RR

  Chromatin was prepared from HeLa cells treated with TNF-α (30ng/ml,1h),cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 15 µg of cross-linked chromatin, 5 µg of NF-κB p65 (80979-1-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR

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  Affinity and Kinetic Characterization of 80979-1-RR

  Biolayer interferometry (BLl) kinetic assays of 80979-1-RR against Human NF-κB p65 were performed. The affinity constant is 3.57 nM.

• Featured Product
• KD/KO Validated

NF-κB p65 Uni-rAb^TM Recombinant Antibody for WB, IF/ICC, FC (Intra), IP, ChIP, ELISA

Cat No. 80979-1-RR

Clone No.4C7

Publications(74)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (1)

Applications

WB, IF/ICC, FC (Intra), IP, ChIP, ELISA

Synonyms

NF κB, p65, p65; RELA, RELA, 4C7

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, MCF-7 cells, MOLT-4 cells, Jurkat cells, Raji cells, NIH/3T3 cells, HSC-T6 cells
• Positive IP detected in: HeLa cells
• Positive IF/ICC detected in: HepG2 cells, TNF alpha treated HT-1080 cells
• Positive FC (Intra) detected in: HepG2 cells
• Positive ChIP detected in: TNF alpha treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:40000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunofluorescence (IF)/ICC: IF/ICC : 1:125-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension
• Chromatin immunoprecipitation (ChIP): CHIP : 1:10-1:100

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 68 publications below
• IF: See 13 publications below

Product Information

80979-1-RR targets NF-κB p65 in WB, IF/ICC, FC (Intra), IP, ChIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, pig
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: NF-κB p65 fusion protein Ag1199
• Full Name: v-rel reticuloendotheliosis viral oncogene homolog A (avian)
• Calculated Molecular Weight: 65 kDa
• Observed Molecular Weight: 65 kDa
• GenBank Accession Number: BC011603
• Gene Symbol: NF-κB p65
• Gene ID (NCBI): 5970
• RRID: AB_2918923
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• 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 sequence-specific DNA-binding protein complex which regulates the expression of viral genomes, including the human immunodeficiency virus, and a variety of cellular genes, particularly those involved in immune and inflammatory responses. The members of the NF-kB family in mammalian cells include the proto-oncogene c-Rel,p50/p105 (NFkB1), p65 (RelA), p52/p100 (NFkB2), and RelB. All of these proteins share a conserved 300-amino acid region known as the Rel homology domain which is responsible for DNA binding, dimerization, and nuclear translocation of NF-kB. The p65 subunit is a major component of NF-kB complexes and is responsible for trans-activation. NF-kB heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kB p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of IkB upon NF-kB 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-kB complex. It associates with chromatin at the NF-kB promoter region via association with DDX1. This antibody is a rabbit polyclonal antibody raised against residues near the N terminus of human RELA.

Publications

Species	Application	Title
mouse	WB	Adv Sci (Weinh); Ccl2-Induced Regulatory T Cells Balance Inflammation Through Macrophage Polarization During Liver Reconstitution; Authors - Rui Wang
mouse	WB	Redox Biol; Deficiency of S100 calcium binding protein A9 attenuates vascular dysfunction in aged mice; Authors - Boying Zhao
mouse		J Exp Clin Cancer Res; METTL3 recruiting M2-type immunosuppressed macrophages by targeting m6A-SNAIL-CXCL2 axis to promote colorectal cancer pulmonary metastasis; Authors - Peng Ouyang
mouse	WB	Sci Total Environ; Effects of chronic low-level lead (Pb) exposure on cognitive function and hippocampal neuronal ferroptosis: An integrative approach using bioinformatics analysis, machine learning, and experimental validation; Authors - Yingsi Cao
pig	WB	Int J Biol Macromol; Angelica sinensis aboveground part polysaccharide and its metabolite 5-MT ameliorate colitis via modulating gut microbiota and TLR4/MyD88/NF-κB pathway; Authors - Yuan-Feng Zou
human	WB	Int J Biol Macromol; A20 ameliorates Aspergillus fumigatus keratitis by promoting autophagy and inhibiting NF-κB signaling; Authors - Fang Han