- 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
- Submit a Review
- Product Citations
- publications
![a. A21012A_PURE_NF-kB-p65_Antibody_WB_020623 a. A21012A_PURE_NF-kB-p65_Antibody_WB_020623](https://d1spbj2x7qk4bg.cloudfront.net/Admin/Public/GetImage.ashx?Image=/Files/Images/media_assets/products/product_images/A21012A_PURE_NF-kB-p65_Antibody_WB_020623-.png&Width=240&Height=300&altFmImage_path=&Compression=90&Crop=5)
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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 -
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 -
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 -
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).
Cat # | Size | Price | Quantity Check Availability | Save | ||
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611351 | 25 µg | £83 | ||||
611352 | 100 µg | £264 |
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 DetailsProduct 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
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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
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- Giridharan S and Srinivasan M. 2018. J Inflamm Res. 11:407-419.
- Ren C, et al. 2021. Cell Death Differ. 29:381-392.
- Yu S, et al. 2020. FEBS Lett. 594:1368-1378.
- Gene ID
- NA
- UniProt
- View information about NF-kB p65 on UniProt.org
Related FAQs
Other Formats
View All NF-κB p65 Reagents Request Custom ConjugationDescription | Clone | Applications |
---|---|---|
Purified anti-NF-κB p65 | A21012A | WB,ICC,IHC-P,ICFC |
Alexa Fluor® 647 anti-NF-κB p65 | A21012A | ICFC |
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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.
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Purified anti-NF-κB p65
Whole cell extracts (15 µg total protein) from the indicated... IHC staining of purified anti-NF- κB p65 (clone A21012A) on ... HeLa cells were fixed with 4% PFA Fixation Buffer (Cat. No. ... HeLa cells were fixed with Fixation Buffer (Cat. No. 420801)... -
Alexa Fluor® 647 anti-NF-κB p65
Peripheral blood lymphocytes were fixed and permeabilized wi...
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