NF-κB p65 Monoclonal antibody

• Featured Product
• KD/KO Validated

NF-κB p65 Monoclonal Antibody for IF, IHC, WB, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

Human and More (3)

Applications

WB, IHC, IF, CoIP, ELISA

Conjugate

Unconjugated

Publications

102

CloneNo.

1B12D11

Cat no : 66535-1-Ig

Synonyms

NF-kB, NFKB3, NF-κB, NF-κB p65, NF-κB3, p65, RELA, Transcription factor p65

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Validation Data Gallery

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  WB analysis of HEK-293 using 66535-1-Ig

  WB result of p65; RELA antibody (66535-1-Ig; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-p65; RELA transfected HEK-293 cells.

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  WB analysis using 66535-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66535-1-Ig (NF-κB p65 antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  IHC staining of human breast cancer using 66535-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66535-1-Ig (p65; RELA antibody) at dilution of 1:300 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66535-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66535-1-Ig (p65; RELA antibody) at dilution of 1:300 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 66535-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using 66535-1-Ig (p65; RELA antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, NIH/3T3 cells, MOLT-4 cells, Jurkat cells, Raji cells
• Positive IHC detected in: human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: HepG2 cells

Planning an IF experiment? We recommend our CoraLite®488 and CoraLite®594 conjugated versions of this antibody.

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:4000
• Immunohistochemistry (IHC): IHC : 1:150-1:600
• Immunofluorescence (IF): IF : 1:50-1:500

Sample-dependent, check data in validation data gallery

Published Applications

• KD/KO: See 1 publications below
• WB: See 90 publications below
• IHC: See 7 publications below
• IF: See 17 publications below
• CoIP: See 1 publications below

Product Information

66535-1-Ig targets NF-κB p65 in WB, IHC, IF, CoIP, ELISA applications and shows reactivity with Human samples.

• Tested Reactivity: Human
• Cited Reactivity: human, bovine, pig
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• 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: RELA
• Gene ID (NCBI): 5970
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• 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.

Background Information

Nuclear factor k B (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-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B 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-kappa-B complex. It associates with chromatin at the NF-kappa-B promoter region via association with DDX1.

Publications

Species	Application	Title
rat		Biochem Pharmacol; Magnesium isoglycyrrhizinate ameliorates fructose-induced podocyte apoptosis through downregulation of miR-193a to increase WT1.; Authors - Tu-Shuai Li
human	IF	Cancer Biol Med; Neutrophil extracellular traps mediate the crosstalk between glioma progression and the tumor microenvironment via the HMGB1/RAGE/IL-8 axis.; Authors - Caijun Zha
mouse	WB	Biomed Res Int; Negative Regulation of Tec Kinase Alleviates LPS-Induced Acute Kidney Injury in Mice via theTLR4/NF-κB Signaling Pathway.; Authors - Wei Zhang
human	WB	Aging (Albany NY); Silencing of lncRNA MIAT alleviates LPS-induced pneumonia via regulating miR-147a/NKAP/NF-κB axis.; Authors - Min Liu
mouse	WB	ACS Chem Neurosci; Quercitrin Rapidly Alleviated Depression-like Behaviors in Lipopolysaccharide-Treated Mice: The Involvement of PI3K/AKT/NF-κB Signaling Suppression and CREB/BDNF Signaling Restoration in the Hippocampus; Authors - Yan Sun
mouse	WB	Med Sci Monit; Live Combined B. subtilis and E. faecium Alleviate Liver Inflammation, Improve Intestinal Barrier Function, and Modulate Gut Microbiota in Mice with Non-Alcoholic Fatty Liver Disease.; Authors - Jie Jiang