NF-κB p65 Rekombinanter Antikörper

• Featured Product
• KD/KO Validated

NF-κB p65 Rekombinant Antikörper für WB, IF/ICC, FC (Intra), IP, ELISA, ChIP-qPCR

Wirt / Isotyp

Kaninchen / IgG

Getestete Reaktivität

human, Maus, Ratte und mehr (1)

Anwendung

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

Konjugation

Unkonjugiert

Publikationen(112)

CloneNo.

4C7

Kat-Nr. : 80979-1-RR

Synonyme

NF κB, p65, p65; RELA, RELA, 4C7

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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 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).

• 

  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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  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 either non-treated or treated with hTNF-α (30 ng/ml, 1 h). 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.

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Geprüfte Anwendungen

• Erfolgreiche Detektion in WB: HeLa-Zellen, HEK-293-Zellen, Jurkat-Zellen, MCF-7-Zellen, MOLT-4-Zellen, NIH/3T3-Zellen, Raji-Zellen
• Erfolgreiche IP: HeLa-Zellen
• Erfolgreiche Detektion in IF/ICC: HepG2-Zellen, TNF alpha treated HT-1080 cells
• Erfolgreiche Detektion in FC (Intra): HepG2-Zellen
• Erfolgreiche Detektion in ChIP-qPCR: hTNF-α-HeLa, 30 ng/ml, 1 h HeLa cells

Empfohlene Verdünnung

• Western Blot (WB): WB : 1:5000-1:40000
• Immunpräzipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunfluoreszenz (IF)/ICC: IF/ICC : 1:125-1:500
• Durchflusszytometrie (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension
• CHImmunpräzipitation (IP)-QPCR: CHIP-QPCR : 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

Veröffentlichte Anwendungen

• WB: See 100 publications below
• IHC: See 1 publications below
• IF: See 22 publications below

Produktinformation

80979-1-RR bindet in WB, IHC, IF/ICC, FC (Intra), IP, ELISA, ChIP-qPCR NF-κB p65 und zeigt Reaktivität mit human, Maus, Ratten

• Getestete Reaktivität: human, Maus, Ratte
• In Publikationen genannte Reaktivität: human, Hausschwein, Maus, Ratte
• Wirt / Isotyp: Kaninchen / IgG
• Klonalität: Rekombinant
• Typ: Antikörper
• Immunogen: NF-κB p65 fusion protein Ag1199
• Vollständiger Name: v-rel reticuloendotheliosis viral oncogene homolog A (avian)
• Berechnetes Molekulargewicht: 65 kDa
• Beobachtetes Molekulargewicht: 65 kDa
• GenBank-Zugangsnummer: BC011603
• Gene symbol: NF-κB p65
• Gene ID (NCBI): 5970
• Konjugation: Unkonjugiert
• Form: Liquid
• Reinigungsmethode: Protein-A-Reinigung
• Lagerungspuffer: PBS with 0.02% sodium azide and 50% glycerol
• Lagerungsbedingungen: Bei -20°C lagern. Nach dem Versand ein Jahr lang stabil Aliquotieren ist bei -20^oC Lagerung nicht notwendig. 20ul Größen enthalten 0,1% BSA.

Hintergrundinformationen

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 monoclonal antibody raised against residues near the N terminus of human RELA.

Publikationen

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	Small; Polymer-Encapsulated Catalase for Targeted Redox Regulation in Acute Liver Injury; Authors - Feifei Li
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	Carbohydr Polym; Structural characterizations and antiaging activities of hydrolyzed low-molecular-weight polysaccharides from Polygoni Multiflori Radix Praeparata; Authors - Wei Xu
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