NF-κB p65 Polyclonal antibody

• Featured Product
• KD/KO Validated

NF-κB p65 Polyclonal Antibody for FC, IF, IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (7)

Applications

WB, IP, IHC, IF, FC, CoIP, ChIP, Cell treatment, ELISA

Conjugate

Unconjugated

Publications

1103

Reviews (7)

Cat no : 10745-1-AP

Synonyms

NF Kappa B, NF κB, NFKB 3, NFKB p65, NFKB3, NF-κB, NF-κB3, p65, RELA, Transcription factor p65

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Validation Data Gallery

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  WB analysis of Hek-293 cells using 10745-1-AP

  WB result of RELA, p65antibody (10745-1-AP, 1:500) with si-Control and si-RELA,p65 transfected HeLa cells.

• 

  WB analysis using 10745-1-AP

  HeLa nuclear fraction and cytoplasmic fraction were subjected to SDS PAGE saparately, followed by western blot with 10745-1-AP (NF-κB p65 antibody) at dilution of 1:6000 incubated at room temperature for 1.5 hours. The membrane was stripped, reblotted with lamin B1 (12987-1-AP) & GAPDH (60004-1-Ig) antibodies as the loading control.

• 

  WB analysis using 10745-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10745-1-AP (NF-κB p65 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Jurkat using 10745-1-AP

  Jurkat cells were subjected to SDS PAGE followed by western blot with 10745-1-AP (p65 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MCF-7 using 10745-1-AP

  MCF7 cells were subjected to SDS PAGE followed by western blot with 10745-1-AP (p65 antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.

• 

  WB analysis of K-562 using 10745-1-AP

  K-562 cells were subjected to SDS PAGE followed by western blot with 10745-1-AP (p65 antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.

• 

  IP experiment of HeLa using 10745-1-AP

  IP Result of anti-p65 (IP:10745-1-AP, 3ug; Detection:10745-1-AP 1:1000) with HeLa cells lysate 5000ug.

• 

  IHC staining of human breast cancer using 10745-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 10745-1-AP (NF-κB p65 antibody) at dilution of 1:200 (under 0x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 10745-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer using 10745-1-AP (p65 antibody) at dilution of 1:100 (under 10x lens).

• 

  IHC staining of human stomach using 10745-1-AP

  Immunohistochemical analysis of paraffin-embedded human stomach using 10745-1-AP (p65 antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human stomach using 10745-1-AP

  Immunohistochemical analysis of paraffin-embedded human stomach using 10745-1-AP (p65 antibody) at dilution of 1:50 (under 40x lens).

• 

  IHC staining of human liver cancer using 10745-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer using 10745-1-AP (p65 antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human liver cancer using 10745-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer using 10745-1-AP (p65 antibody) at dilution of 1:50 (under 40x lens).

• 

  IF Staining of HepG2 using 10745-1-AP

  Immunofluorescent analysis of (-20℃ Ethanol) fixed HepG2 cells using 10745-1-AP (p65; RELA antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  FC experiment of HeLa using 10745-1-AP

  1X10^6 HeLa cells were stained with 0.2ug p65; RELA antibody (10745-1-AP, red) and control antibody (blue). Fixed with 90% MeOH blocked with 3% BSA (30 min). Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) with dilution 1:1000.

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Tested Applications

• Positive WB detected in: A431 cells, Hek-293 cells, Jurkat cells, MCF-7 cells, K-562 cells, HeLa cells, Raji cells, NIH/3T3 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human breast cancer tissue, human stomach tissue, human liver 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
• Positive FC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for IP and 1:500-1:2000 for WB
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF): IF : 1:50-1:500

Sample-dependent, check data in validation data gallery

Published Applications

• KD/KO: See 20 publications below
• WB: See 995 publications below
• IHC: See 85 publications below
• IF: See 169 publications below
• IP: See 10 publications below
• ELISA: See 1 publications below
• CoIP: See 6 publications below
• ChIP: See 5 publications below

Product Information

10745-1-AP targets NF-κB p65 in WB, IP, IHC, IF, FC, CoIP, ChIP, Cell treatment, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, goat, CHICKEN, monkey, rabbit, fish, hamster, pig
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• 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: Antigen affinity 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. 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-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	IF	Blood; PMID: 32291445; Authors - Kunpeng Wu
human	WB	Nat Microbiol; IFI16 directly senses viral RNA and enhances RIG-I transcription and activation to restrict influenza virus infection.; Authors - Zhimin Jiang
human	WB	Sci Adv; JLP-centrosome is essential for the microtubule-mediated nucleocytoplasmic transport induced by extracellular stimuli.; Authors - Clement M Lee
human	WB,IF	Nucleic Acids Res; p38-mediated FOXN3 phosphorylation modulates lung inflammation and injury through the NF-κB signaling pathway; Authors - Xinxing Zhu
mouse	WB	Nat Commun; REV-ERBα integrates colon clock with experimental colitis through regulation of NF-κB/NLRP3 axis.; Authors - Shuai Wang
mouse	WB	Nat Commun; Reducing Akt2 in retinal pigment epithelial cells causes a compensatory increase in Akt1 and attenuates diabetic retinopathy; Authors - Haitao Liu

Reviews

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

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FH

Maximilian (Verified Customer) (06-30-2023)

Quite a bit of unspecific binding. Did not work in mouse cell lines.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Mainly Human Breast Cancer Cell lines (MDA-MB-436, MDA-MB-468, MDA-MB-231)

FH

Xie (Verified Customer) (09-29-2022)

NF-κB p65 Antibody staining.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:250
• Cell Tissue Type: hela

NF-κB p65 Antibody Immunofluorescence validation (1:250 dilution) in hela (Cat no:10745-1-AP)

FH

Murali (Verified Customer) (01-24-2022)

Very nice antibody for IHC

• Applications: Immunohistochemistry
• Primary Antibody Dilution: 1 to 200
• Cell Tissue Type: pharyngeal mucosal

NF-κB p65 Antibody Immunohistochemistry validation (1 to 200 dilution) in pharyngeal mucosal (Cat no:10745-1-AP)

FH

Azita (Verified Customer) (05-31-2021)

NSC-34 cells (motor neuron-like cells) strong labelling in WB

• Applications: Western Blot
• Primary Antibody Dilution: 1/2000
• Cell Tissue Type: Human spinal cord (cervical) tissue

FH

Zehua (Verified Customer) (12-17-2019)

After siRNA treatment for 48 hours, specific signal shown here. Works excellent!

• Applications: Western Blot, Cell culture,
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: K562

NF-κB p65 Antibody Western Blot,Cell culture, validation (1:1000 dilution) in K562 (Cat no:10745-1-AP)

FH

Feng-qian (Verified Customer) (12-07-2018)

• Applications: Western Blot, Immunofluorescence,
• Primary Antibody Dilution: 1/500
• Cell Tissue Type: HaCaT

FH

Fraser (Verified Customer) (06-06-2018)

• Applications: Other (please state below),

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