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Purified anti-NF-κB p65 Antibody

Pricing & Availability
Clone
A21012A (See other available formats)
Regulatory Status
RUO
Other Names
RelA, P65, NFKB3, Nuclear Factor NF-Kappa-B P65 subunit, Transcription Factor P65, NF-Kappa-B Transcription Factor P65, RELA Proto-Oncogene NF-KB subunit
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
a. A21012A_PURE_NF-kB-p65_Antibody_WB_020623
Whole cell extracts (15 µg total protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL purified anti-NF- κB p65 (clone A21012A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-GAPDH (Cat. No. 607904) was used as a loading control at a 1:50000 dilution. Lane M: Molecular weight marker
  • a. A21012A_PURE_NF-kB-p65_Antibody_WB_020623
    Whole cell extracts (15 µg total protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL purified anti-NF- κB p65 (clone A21012A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-GAPDH (Cat. No. 607904) was used as a loading control at a 1:50000 dilution. Lane M: Molecular weight marker
  • b. A21012A_PURE_NF-kB-p65_Antibody_IHC-P_020623
    IHC staining of purified anti-NF- κB p65 (clone A21012A) on formalin-fixed paraffin-embedded human colon tissue. Following antigen retrieval using Tris EDTA, the tissue was then incubated without (panel A) or with (panel B) anti-NF- κB p65 (clone A21012A), followed by incubation with 10 µg/mL of Alexa Fluor® 647 goat anti-mouse IgG (Cat. No. 405322) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 60X objective. Scale bar: 50 µm
  • c. A21012A_PURE_NF-kB-p65_Antibody_ICC_020623
    HeLa cells were fixed with 4% PFA Fixation Buffer (Cat. No. 420801), permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with 5.0 µg/mL purified mouse IgG1, κ isotype control (Cat. No. 400102) (panel A) or with 5.0 µg/mL of purified anti-NF- κB p65 (clone A21012A) (panel B), followed by incubation with 2.5 µg/mL of Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 60X objective. Scale bar: 50 µm
  • d. A21012A_PURE_NF-kB-p65_Antibody_ICFC_020623
    HeLa cells were fixed with Fixation Buffer (Cat. No. 420801), permeabilized using True-Phos™ Perm Buffer (Cat. No. 425401), and intracellularly stained with anti-NF- κB p65 (clone A21012A) (filled histogram) or purified Mouse IgG1, κ isotype control (Cat. No. 400102) (open histogram) followed by PE goat anti-mouse IgG (Cat. No. 405307).
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611351 25 µg 120 CHF
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611352 100 µg 385 CHF
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Description

NF-κB p65, also known as NFKB3 and RelA, is one of five members of the nuclear factor kB (NF-kB)/Rel family. Inactive NF-κB p65 is sequestered in the cytoplasm as a complex with IκB inhibitory proteins. Upon activation of the NF-κB pathway, IκB proteins are degraded, thus releasing NF-κB p65 to the nucleus. Binding of NF-κB p65 to transcriptional cofactors is induced via phosphorylation on serines 276, 529, 536, and 471. As a transcription factor, NF-κB p65 is vital to cell proliferation, immune response, survival, and apoptosis. Through regulation of chondrocyte and osteoblast differentiation and survival, NF-κB p65 has a crucial role in skeletal development. Overexpressed in several cancers, NF-κB p65 functions as an oncogene and has been implicated in tumorigenesis, metastasis, tumor angiogenesis, and chemoresistance.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide corresponding to human NF-κB p65
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
IHC-P, ICC, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.25 - 0.50 µg/mL. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1.0 - 10.0 µg/mL is suggested. For immunocytochemistry, a concentration range of 1.25 - 5.0 μg/mL is recommended. For flow cytometric staining, the suggested use of this reagent is ≤ 0.125 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application

Application Notes

When testing by ICC, we recommend fixation with 4% PFA followed by permeabilization with Triton X-100 or ice-cold methanol.

RRID
AB_2936734 (BioLegend Cat. No. 611351)
AB_2936734 (BioLegend Cat. No. 611352)

Antigen Details

Structure
NF-κB p65 is a 551 amino acid protein with a predicted molecular weight of 60 kD.
Distribution

Ubiquitously expressed/ Cytoplasm and Nucleus

Function
Transcription, RAS pathway, host-virus interaction, oncogene
Interaction
NF-kappa-B, TLE5, TLE1, TP53BP2, GSK3B, MAP3K14, HDAC3, AKIP1, FOXP3
Biology Area
Cell Biology, Immuno-Oncology, Immunology, Innate Immunity, Transcription Factors
Molecular Family
Phospho-Proteins
Antigen References
  1. Giridharan S and Srinivasan M. 2018. J Inflamm Res. 11:407-419.
  2. Ren C, et al. 2021. Cell Death Differ. 29:381-392.
  3. Yu S, et al. 2020. FEBS Lett. 594:1368-1378.
Gene ID
NA
UniProt
View information about NF-kB p65 on UniProt.org
Go To Top Version: 1    Revision Date: 02.06.2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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