NF-κB p65 Monoclonal antibody

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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. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

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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 (4% PFA) fixed HepG2 cells using NF-κB p65 antibody (66535-1-Ig, Clone: 1B12D11 ) at dilution of 1:1000 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L), CL594-phalloidin (red).

• Featured Product
• KD/KO Validated

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

Cat No. 66535-1-Ig

Clone No.1B12D11

Publications(294)

Reviews (2)

Host / Isotype

Mouse / IgG1

Reactivity

Human and More (5)

Applications

WB, IHC, IF, CoIP, ChIP, RIP, ELISA

Synonyms

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

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 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

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:4000
• Immunohistochemistry (IHC): IHC : 1:150-1:600

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

• KD/KO: See 3 publications below
• WB: See 258 publications below
• IHC: See 18 publications below
• IF: See 42 publications below
• CoIP: See 2 publications below
• ChIP: See 3 publications below
• RIP: See 1 publications below

Product Information

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

• Tested Reactivity: Human
• Cited Reactivity: human, pig, chicken, bovine, hamster
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag1199

  Product name: Recombinant human p65; RELA protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 1-220 aa of BC011603

  Sequence: MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQ

• 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_2881898
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G 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 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

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
	WB	Brain Behav Immun; Egln3 expression in microglia enhances the neuroinflammatory responses in Alzheimer's disease; Authors - Jiaxin Guan
human	ChIP	Adv Sci (Weinh); Elevated SPARC Disrupts the Intestinal Barrier Integrity in Crohn's Disease by Interacting with OTUD4 and Activating the MYD88/NF-κB Pathway; Authors - Jiayu Wang
	WB	Adv Healthc Mater; Colon-Targeted Ginseng Polysaccharides-Based Microspheres for Improving Ulcerative Colitis via Anti-Inflammation and Gut Microbiota Modulation; Authors - De-Yang Huo
	WB	Cell Rep; METTL3-mediated m6A mRNA methylation regulates neutrophil activation through targeting TLR4 signaling; Authors - Shuhua Luo
	WB	Theranostics; Prussian blue nanozyme-mediated nanoscavenger ameliorates acute pancreatitis via inhibiting TLRs/NF-κB signaling pathway.; Authors - Xue Xie
human	WB	Int J Biol Macromol; SUMOylation of GMFB regulates its stability and function in retinal pigment epithelial cells under hyperglycemia; Authors - Wan Sun

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Udesh (Verified Customer) (04-19-2023)

Lowest membrane part has NF-kB p65 in U2OS cells

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: U2OS

NF-κB p65 Antibody Western Blot validation (1:1000 dilution) in U2OS (Cat no:66535-1-Ig)

FH

Federica (Verified Customer) (01-16-2023)

The band comes out well even when low total protein concentration is loaded (15 ug).

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Dendritic cells